TWI649319B - Bicyclic heterocyclic compounds and therapeutic uses thereof - Google Patents

Abstract

The present invention relates to a novel bicyclic heterocyclic compound, a pharmaceutical composition comprising the compound, and the use of the compound for treating a disease such as cancer.

Description

Bicyclic heterocyclic compound and its therapeutic use

The present invention relates to a novel bicyclic heterocyclic compound, a pharmaceutical composition comprising the compound, and the use of the compound for treating a disease such as cancer.

IAP family

The inhibitor of apoptosis protein (IAP) family includes eight members: XIAP, cIAP1, cIAP2, NAIP, ILP2, ML-IAP, survivin, and BRUCE (also known as apollon). Although the exact mechanism of all eight members of the IAP family is not fully understood, it is known that members of the IAP family can inhibit planned cell death through the ability to directly inhibit members of the caspase family of apoptotic enzymes . The common structural feature of all members of the IAP family is a zinc-bound folding unit containing about 70 amino acids with one to three repeats, also known as the baculovirus IAP repeat (BIR) region.

Many of the interactions between IAP and other proteins are through surface grooves in the BIR region. BIR regions can be classified according to their peptide binding specificity. Among them, there are three types of BIR regions: Type III region, which passes through the proline at the third position (P3), and can specifically excrete with caspase (and similar to caspase) Peptide chain binding (e.g., XIAP BIR3); type II regions, which are similar to type III regions but do not necessarily have proline (e.g. XIAP BIR2); and type I regions, which do not interact with caspase or It binds similarly to a peptide chain (eg, XIAP BIR1) (Eckelman et al. Cell Death and Differentiation 2008; 15: 920-928). The BIR system is a small Zn-coordination region (about 70 amino acids), and a variety of protein systems use its N-terminus to interact with the BIR region groove. BIR antagonists (antagonist) can prevent caspase from binding to BIR, and then increase caspase activity, which leads to auto-ubiquitination and proteasome degradation of IAP.

IAP is overexpressed in many cancers, including kidney cancer, melanoma, colorectal cancer, lung cancer, breast cancer, ovarian cancer, and prostate cancer (Tamm et al., Clin. Cancer Research 2000; 6 (5): 1796-803), It is also associated with tumor growth, pathogenesis, and resistance to chemotherapy and radiation therapy (Tamm 2000).

XIAP

XIAP is a 57 kDa protein with three BIR regions (the second and third BIR regions are bound to caspase) and a RING-type zinc finger (E3 ligase). XIAP also binds to a variety of proteins other than caspase, including ligase receptors such as TAK1 and cofactor TAB1, and MURR1 related to copper homeostasis (Burstein et al., EMBO 2004; 23: 244-254 ), Such as endogenous inhibitors of the second mitochondrial-derived caspase promoter (SMAC), and proteins such as MAGE-D1, NRAGE, etc. whose functions are unknown (Jordan et al., J. Biol. Chem. 2001; 276: 39985-39989).

The BIR3 region binds and inhibits caspase-9, which is an apical caspase in the mitochondrial pathway of caspase activation. The surface groove of the BIR3 region interacts with the N-terminus of the caspase-9 minor unit, rendering the activation site of caspase-9 ineffective and existing as an inactive monomer (Shiozaki et al., Mol. Cell 2003; 11: 519-527).

In addition to caspase binding, XIAP also inhibits apoptosis through other mechanisms. XIAP will form a complex with TAK1 kinase and its co-factor TAB1, and activate the JNK and MAPK signaling pathways, resulting in NF-κB activation (Sanna et al., Mol Cell Biol 2002; 22: 1754-1766) . XIAP also activates NFκB by promoting the translocation of NF-κB to the nucleus and promoting the degradation of IκB (Hofer-Warbinek et al., J. Biol. Chem. 2000; 275: 22064-22068, Levkau et al. , Circ. Res. 2001; 88: 282-290).

Transfection of cells with XIAP can inhibit planned cell death from various apoptotic stimuli (Duckett et al., EMBO 1996; 15: 2685-2694, Duckett et al., MCB 1998; 18: 608-615, Bratton, Lewis, Butterworth, Duckett and Cohen, Cell Death and Differentiation 2002; 9: 881-892).

XIAP is ubiquitinated in all normal tissues, but has improved pathologically in many acute and chronic leukemias, prostate cancer, lung cancer, kidney cancer, and other cancers (Byrd et al., 2002; Ferreira et al., 2001 Hofmann et al., 2002; Krajewska et al., 2003; Schimmer et al., 2003; Tamm et al., 2000). In primary acute myelogenous leukemia (AML), the XIAP expression line is associated with French-American-British (FAB) subtype M4 / M5 (P <0.05) and is associated with AML bud cells (blast). The expression of monocyte markers was correlated. In addition, XIAP was also found to be overexpressed in normal monocytes, but this phenomenon was not seen in granular cells. In AML, the performance of XIAP in patients with more favorable cytogenetics was significantly lower than that in patients with moderate or poor cytogenetics (n = 74; P <0.05) (Tamm et al., Hematol. J. 2004; 5 (6): 489-95).

Overexpression can cause cells to become resistant to multi-agent therapies and lead to poor clinical treatment of diseases, including AML, kidney cancer, melanoma (Tamm et al., Clin. Cancer Research 2000; 6: 1796-1803) and lung cancer (Hofmann et al., J. Cancer Res. Clin. Oncology 2002; 128 (10): 554-60).

XIAP is transcribed by a cap-independent mechanism of transcription initiation, which is performed by a unique internal ribosome entry site (IRES) sequence unit in the 5 'non-transcribed region. When protein synthesis is inhibited in most cells, this mechanism enables XIAP mRNA to activate transcription during cellular stress. The up-regulated response to XIAP transcription produced by cellular stress increases resistance to radiation that induces cell death (Holcik et al., Oncogene 2000; 19: 4174-4177).

The XIAP inhibitory mechanism has been studied in vitro by several technologies, including RAN gene silencing test, gene knock-out test, peptide ligand simulation test, and small molecule antagonists. It has been proven that monotherapy can help apoptotic diseases to promote multiple tumors. Sensitivity to chemotherapy, including bladder cancer (Kunze et al., 2008; 28 (4B): 2259-63). XIAP knockout mice can be born at the expected Mendelian frequency without significant physiological or tissue defects, and have a normal lifespan (Harlin et al., Mol. Cell Biol. 2001; 21 (10 ): 3604-3608). This result shows that when the lack of XIAP activity does not cause toxicity to normal tissues, it can be used as a therapeutic window for tumor cells. More in-depth research shows that XIAP has a critical distinction between hepatocyte type 1 and type 2 apoptosis, so it should be used with caution in patients with background of liver symptoms (Jost et al., Nature, 2009, 460, 1035-1041). It has been pointed out that in XIAP knockout mice, the amounts of cIAP1 and cIAP2 are up-regulated, and the compensation mechanism can be used to protect the lesion, so pan-inhibition may be necessary for functional knockout . Similarly, cIAP1 and cIAP2 knockout mice were asymptomatic (Conze et al., Mol. Biol. Cell 2005; 25 (8): 3348-56). Although the absence of any of the IAPs did not cause significant symptoms in mice, deletion of cIAP1 and cIAP2 or XIAP resulted in intermediate embryo death (Moulin, EMBO J., 2012).

For example, SMAC's endogenous IAP antagonists have been used as targets for this family of therapeutic agents. SMAC peptides sensitize tumor cells to chemotherapy. When SMAC peptides are used in combination with platinum-containing drugs and tumor-necrosis factor alpha-related apoptosis-inducing ligands (TRAIL), it can delay tumor cell growth (Fulda et al. , Nat. Med. 2002; 808-815; Yang et al., Cancer Res. 2003; 63: 831-837).

The natural substance of the letterbell quinone (embellin) is considered to be able to bind to the surface groove of the BIR3 region of XIAP, and its binding affinity is similar to that of the natural SMAC peptide. Letterbox quinone can cause apoptosis of cell lines in vitro and delay tumor growth of xenograft (Nikolovska-Coleska et al., J. Med. Chem. 2004; 47 (10): 2430-2440; Chitra et al. ,, Chemotherapy 1994; 40: 109-113).

XIAP antisense oligonucleotides have been developed as therapeutic agents for the treatment of solid tumors and hematological malignancies. In in vitro tests, these anti-strand oligonucleotides can block approximately 70% of protein expression, cause apoptosis, make cells sensitive to chemotherapy, and delay tumor growth in in vivo tests. One of these agents, AEG351156, has been clinically tested (Hu et al., Clin. Cancer Res. 2003; 9: 2826-2836; Cummings et al., Br. J. Cancer 2005; 92: 532- 538).

XIAP antagonists that have been developed include peptidomimetics and synthetic agents. The peptidomimetic target BIR3 region mimics the SMAC blockade of caspase-9 binding to XIAP, and has been proven to induce apoptosis in various tumor cell lines in the form of a single agent, and can also be used as chemosensitization Agents and are currently undergoing clinical trials (Oost et al., J. Med. Chem. 2004; 47: 4417-4426; Sun et al., Bioorg. Med. Chem. Lett. 2005; 15: 793-797) .

The synthetic small molecule antagonists of the BIR3 and BIR2 regions also have anti-tumor effects in various models, including apoptosis-inducing experiments with annexin-V staining, and IC50 of more than one-third in the test plate of NCI60 cell line is <10 μM. XIAP antagonists also caused meter-related cell death in primary cultured leukemia cells, cell death in five of the five chronic lymphocytic white blood cell lines, and cell death was observed in four of the five acute myeloid white blood cell lines ( Schimmer et al., Cancer Cell 2004; 5: 25-35; Berezovskaya et al., Cancer Res. 2005; 65 (6): 2378-86).

The quality of XIAP protein in tumor cell lines is quite high, and it has a negative correlation with the sensitivity of some anticancer drugs, especially cytarabine and other nucleoside drugs (Tamm et al., Clin. Cancer Research 2000; 6 : 1796-1803). In two preclinical models of pancreatic cancer in vivo, XIAP inhibition may be related to TRAIL-induced anticancer activity (Vogler 2008). Gene expression and transfection studies have shown that the increase in the expression of the apoptosis inhibitor XIAP is related to anoikis resistance and also related to the survival of human circulating prostate cancer cells, leading to cancer metastasis. Small molecule antagonists have been found to achieve anti-metastatic effects in these models (Berezovskaya et al., Cancer Res. 2005; 65 (6): 2378-86).

XIAP has also been linked to other pathways related to cancer and other diseases, and this finding is beneficial to the development of XIAP target agents. The E3 ligase activity of the RING finger region of XIAP can bind to the upstream regions of TAB1 and BMP receptors (type 1), indicating that XIAP can transmit information through the TGF-β-mediated pathway (Yamaguchi et al ., EMBO 1999; 179-187). Excessive expression of focal adhesion kinase (FAK) leads to up-regulation of XIAP performance (Sonoda et al., J. Biol. Chem. 2000; 275: 16309-16315). E3 ligase can be used as a therapeutic target, and molecules that can target E3 ligase activity in other proteins (such as MDM2) have been developed (Vassilev et al., Science 2004; 303: 844-848). Direct or indirect inhibition of XIAP ligase activity can also be effective in treating cancer or other diseases. Dysregulation of apoptotic message transmission may be due to inhibition of IAP function that controls planned cell death and is associated with a variety of diseases, including cell aggregation (e.g., cancer, autoimmunity, inflammatory response, and restenosis (restenosis) or disorders, diseases that cause cell loss due to excessive apoptosis (e.g., stroke, heart failure, neurodegeneration (e.g., Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic spinal cord) Lateral sclerosis), AIDS, ischemia (stroke, myocardial infarction) and osteoporosis).

XIAP is an important apoptosis regulator in experimental autoimmune encephalomyelitis and a potential target drug for the treatment of autoimmune diseases such as multiple sclerosis (MS) (Moore et al., 2004; 203 (1) : 79-93). In the MS animal model, antisense-mediated elimination of XIAP makes paralyzed animals stand up, suggesting that targeting XIAP and other possible IAPs can be used in MS treatment (Hebb et al., Curr. Drug Disc. Tech 2008; 5 (1): 75-7).

cIAP1, cIAP-2, XIAP, and survivin are overexpressed in malignant pleural mesothelioma, and are extremely related to the resistance of artificially cultured mesothelioma cells to cisplatin. The amount of circulating TNF-α in mesothelioma patients before surgery was significantly higher than after surgery. TNF-α causes increased mRNA and protein mass of IAP-1, IAP-2, and XIAP (Gordon et al., 2007). For the inflammatory response caused by exposure to asbestos fibers, the upregulation of NF-κB is highly related to the survival of mesothelioma (Sartore-Bianchi et al., 2007). IAP antagonists have the potential to reverse regulate the pro-survival effect of TNF-α.

Once cIAP1 and cIAP2 decrease, the ability of cell lines to regulate the expression of TNF-α is sufficient to kill cells in an autocrine response and this phenomenon is considered to be very important for IAP activity (Nature Reviews Cancer (2010), 10 (8 ), 561-74, Gryd-Hansen, M). However, in vivo experiments, certain tumor types will be surrounded by a network of pro-inflammatory cytokines, so when cIAP1 / 2 decreases, tumor cells will switch to apoptosis by killing the cells, and can pass through TNF-α (or other Lethal receptor cytokine antagonists) initiate apoptosis, in which TNF-α (or other lethal receptor cytokine antagonists) is in the tumor microenvironment by surrounding cells (such as tumor-associated macrophages) or the tumor cells themselves generate. Specific tumor types such as breast cancer, ovarian cancer, and melanoma show the aforementioned "inflammatory phenotype", which has the potential to be a target for IAP antagonists.

cIAP1 and cIAP2

Cell IAP (cIAP) 1 and 2 are closely related members of the IAP family, with three BIR regions, a RING region, and a caspase recruitment (CARD) region. Functional nuclear export signals are present in the CARD region of cIAP1, which is highly correlated with cell differentiation (Plenchette et al., Blood 2004; 104: 2035-2043). This CARD region is specifically found in cIAP1 and cIAP2 in the IAP family of proteins. These two gene lines exist on the 11q22 chromosome, and their high genetic similarity is considered to be caused by gene replication.

Like XIAP and survivin, cIAP1 is widely expressed in tumor cell lines, and is particularly highly expressed in colorectal cancer, pneumonia, ovarian cancer, kidney cancer, CNS, and breast cancer (Tamm et al., Clin. Cancer Res. 2000; 6: 1796-1803). The performance of cIAP2 is generally more restricted, and it is considered to be regulated by the constitutive ubiquitination and degradation of cIAP1 (Conze et al., Mol. Biol. Cell 2005; 25 (8): 3348-56; Mahoney et al., PNAS 2008; 105: 11778-11783). Immune tissue analysis and western blot analysis used to determine whether cIAP1 and cIAP2 are potential oncogenes. In a variety of lung cancers, cIAP1 and cIAP2 are overexpressed with high or low gene copy numbers (Dia et al., Human Mol Genetics 2003; 12 (7): 791-801). The expression of cIAP1 appears to be highly correlated with precancerous adenocarcinoma (Hofmann et al., J. Cancer Res. Clin. Oncology 2002; 128 (10): 554-60).

Like XIAP, the increase in cIAP1 and cIAP2 expression and the decrease in endogenous inhibitor expression are related to drug resistance. In vitro tests have shown that cIAP overexpression correlates with resistance to DNA alkylating agents such as carboplatin, cisplatin, and topoisomerase inhibitor VP-16 (Tamm et al ., Clin. Cancer Res. 2000; 6: 1796-1803). After cisplatin and doxorubicin treatment, more cIAP1 and survivin were found in thyroid cancer cells. The resistance of cells to chemotherapeutic agents such as paclitaxel will cause reduced SMAC performance and cause mitochondria to secrete a small amount of SMAC protein. The down-regulation of cIAP1 and survivin can increase the cytotoxicity of cisplatin and doxorubicin, but overexpression of SMAC can improve the efficacy of paclitaxel. However, cIAP1 and survivin gene silencing caused by RNA interference will restore the drug sensitivity of cisplatin and doxorubicin (Tirrò et al .; Cancer Res. 2006; 66 (8): 4263-72).

For example, the SMAC mimic of LBW242 was initially considered as the main target of XIAP. However, studies have shown that cIAP1 can cause cell degradation through autologous ubiquitination in cells (Yang et al., J. Biol. Chem. 2004; 279 (17): 16963-16970) and may be related to apoptosis. cIAP1 and tumor necrosis factor (TNF) -α-induced (or stimulated) siRNAs have been found to have synergistically, making cell lines more drug-sensitive (Gaither et al. Cancer Res. 2007; 67 (24 ): 11493-11498).

cIAP1 and cIAP2 are important regulators of the NF-κB signaling pathway. They participate in a variety of biological programs, especially innate and acquired immune responses, and cell proliferation and survival. Deregulation of the NF-κB pathway is related to inflammation and cancer, including hepatitis and ulcerative colitis, gastritis, hepatocellular carcinoma, colorectal and gastric cancer, and angiogenesis and cancer metastasis (Shen et al. ,, Apoptosis 2009; 14: 348-363).

On ligand binding, the TNF receptor (TNFR) recruits TNFR-related death regions (TRADD) and receptor-interacting protein (RIP) 1. Then, TRAF2 and cIAP1 / cIAP2 were recruited to form a large cell membrane complex. After RIP1 is ubiquitinated, these polyubiquitin chains serve as downstream kinase binding sites, resulting in the NF-κB pathway message transmission effect (Ea et al., Mol. Cell 2006; 22: 245-257; Wu et al. ., Nat. Cell Biol. 2006; 8: 398-406). Although the derivatization is quite complex and not fully understood, cIAP1 and cIAP2 are considered to be key factors for TNF-α-mediated regulation of NF-κB signaling and ligand-independent / typical NF-κB signaling (Varfolomeev et al., Cell 2007; 131 (4): 669-81). cIAP1 and cIAP2 can be combined with TRAF2. Among them, TRAF2 is an adapter protein in the typical and interactive NF-κB signaling pathway and the MAPK signaling pathway (Rothe et al., Cell 2005; 83: 1243-1252). In in vitro experiments, cIAP1 and cIAP2 can directly target RIP1 for ubiquitination (Betrand et al., Mol. Cell 2008; 30: 689-700).

TNF-α regulates many cell functions, including apoptosis, inflammatory response, immune response, and cell growth and differentiation (Trace et al., Annu. Rev. Med. 1994; 45: 491-503), and therapeutic IAP antagonists It is effective when these cell functions are affected.

Many malignant tumors produce TNF-α, which is one of the key drivers of cancer-related inflammation, which drives tumor development and / or progression. cIAPs protect cancer cells from the lethal effects of TNF-α.

NAIP

NAIP is the first IAP to be discovered (Roy et al., Cell 1995; 80: 167-178). NAIP is quite special in IAP, because it has nucleotide binding and oligomerization regions and is rich in leucine repeats. This leucine-rich repeats are rich in light with those in common innate immune-related proteins. The amino acid repeats are similar. Currently, NAIP is also overexpressed in some cancers including breast cancer and esophageal cancer (Nemoto et al., Exp. Mol. Pathol. 2004; 76 (3): 253-9), and it is also overexpressed in MS ( Choi et al., J. Korean Med. 2007; 22 Suppl: S17-23; Hebb et al., Mult. Sclerosis 2008; 14 (5): 577-94).

ML-IAP

Melanoma apoptosis inhibitory protein (ML-IAP) contains a single BIR and RING finger structure. In apoptosis induced by death receptors and chemotherapeutic agents, ML-IAP is a very effective inhibitor and can be used as a direct inhibitor of downstream effector caspase (Vucic et al., Curr. Biol. 2000; 10 (21): 1359-66). ML-IAP is also known as baculovirus IAP repeat-containing protein 7 (BIRC7), renal apoptosis inhibitory protein (KIAP), RING finger protein 50 (RNF50), and apoptosis inhibitor (Livin). The BIR region of ML-IAP has an evolutionarily conserved fold that is essential for anti-apoptotic activity. It has been found that most melanoma cell lines show high levels of ML-IAP, but the ML-IAP performance of primary black cells is almost undetectable. Melanoma cells are more significantly resistant to drug-induced apoptosis. Increased expression of ML-IAP may cause resistance of melanoma cells to apoptosis stimuli, and is potentially related to the pathogenesis of malignant tumors.

ILP2

ILP-2, also known as BIRC8, has a single BIR region and a RING region. ILP-2 is expressed only in normal cells of testes and binds to caspase-9 (Richter et al, Mol. Cell. Biol. 2001; 21: 4292-301).

Survivin

Survivin, also known as BIRC5, inhibits caspase 3 and caspase 7, but its main function is the regulation of cell mitosis rather than the regulation of apoptosis. Survivin helps microtubule production in mitotic spindles and is involved in apoptosis in the cell cycle. Survivin apoptosis is inhibited in colorectal cancer (Kawasaki et al., Cancer Res. 1998; 58 (22): 5071-5074) and stage III gastric cancer (Song et al., Japanese J. Clin. Oncol. 2009 39 (5): 290-296).

BRUCE

BRUCE (ubiquitinated binding enzyme with BIR repeats) is a trans-Golgi network surrounding membrane protein, which has a single BIR region and is most similar to survivin. BRUCE can be inhibited by three mechanisms: (i) SMAC binding, (ii) HtrA2 protease, and (iii) caspase-mediated cleavage. In addition, BRUCE can also serve as an E2 / E3 ubiquitin ligase through its ubiquitinated binding (UBC) region.

The present invention provides a compound of formula (I). The present invention provides compounds useful in therapy, particularly for cancer treatment. Compounds of formula (I) can be antagonists of the IAP protein family, especially antagonists of XIAP and / or cIAP (such as cIAP1 and / or cIAP2), and can be used to treat IAP-mediated abnormal physical conditions.

According to a first aspect of the present invention, a compound of formula (I) is provided: (I) or a tautomer thereof, a stereochemical isomer thereof, a pharmaceutically acceptable salt thereof, or a solvate thereof; wherein X is CR ⁴ , N or NR ³ ; wherein when X is CR ⁴ , Then U is nitrogen and R ⁶ is oxo; or when X is N, U is carbon and R ⁶ is methylol or -CH (OR ^x ) CH ₂ OR ^z ; or when X is NR ³ , then U represents carbon and R ⁶ represents pendant oxygen; the dashed bond (-------) represents a single or double bond, wherein at least two of the dashed bonds represent a double bond; R ¹ and R ² independently represent hydrogen or methyl; R ³ represents hydrogen, methyl or -NH ₂ ; R ⁴ represents hydrogen, methyl, hydroxymethyl, -NH ₂ or fluorine; R ⁵ represents unsubstituted ortho butyl, or one or two of fluoro-substituted benzyl group on the phenyl; and R ^x and R ^z independently represent hydrogen or methyl lines.

In another aspect of the invention, a compound of formula (I) is provided for use in the prevention or treatment of a disease or symptom as described herein, a pharmaceutical composition comprising a compound of formula (I), and a compound of formula (I) Methods.

definition

Unless otherwise specified in the specification, the chemical formula (I) (including the use, method or other aspects of the invention) referred to in all sections of the present specification includes all other sub-formulas defined herein. ), Sub-group, preference, examples and experimental examples.

"IAP" means any of the members of the IAP family XIAP, cIAP (cIAP1 and / or cIAP2), NAIP, ILP2, ML-IAP, survivin and / or BRUCE, preferably XIAP, cIAP1, cIAP2, ML- IAP, more preferably XIAP, cIAP1 and / or cIAP2, and most preferably XIAP and / or cIAP1. In particular, it means the BIR region of IAP, preferably the BIR region of XIAP, cIAP1 or cIAP2.

"One or more members of the IAP family" refers to any one of the members of the IAP family, preferably XIAP, cIAP1, and / or cIAP2, and more preferably XIAP and / or cIAP1.

"Pharmaceutical effect" refers to a measurement of the activity of a drug, which is expressed in an amount capable of producing a predetermined strength effect. Stronger drugs can trigger larger reactions at low concentrations. Pharmacodynamics is proportional to affinity and efficacy. Affinity refers to the ability of a drug to bind to a receptor. Efficacy is the relationship between receptor occupancy and the ability to trigger a molecular, cell, tissue or system response.

"Antagonist" refers to a type of receptor ligand or drug that can block or inhibit a synergist-mediated biological response. Antagonists have affinity for their cognate receptors, but they do not have a potent effect on their cognate receptors, and after binding, they block any ligands on the receptor (such as endogenous ligands or receptors , Agonists or reactivators), and block their function. Antagonism can be caused directly or indirectly, and can be mediated by any mechanism at any physiological stage. An example of indirect antagonism is indirect antagonism of cIAP, causing cIAP to be ubiquitinated and degraded. Therefore, the antagonistic action of the ligand can be performed in different ways in different environments. Antagonists bind to the active site or allosteric site of the receptor to produce an antagonistic effect, or they can interact with specific binding sites that are not generally related to the physiological regulation of receptor activity. Antagonist activity can be reversible or irreversible, depending on the duration of the antagonist-receptor complex, and this is related to the antagonist-receptor binding characteristics.

As used herein, "treatment" refers to the treatment of abnormal physical conditions (such as symptoms, abnormalities or diseases), whether directed at humans or animals (i.e. veterinary applications), and is generally related to treatment or therapy, in which Desired therapeutic effects, such as inhibiting the development of abnormal physical conditions, including slowing the rate of disease development, pausing the rate of disease development, improving abnormal physical conditions, eliminating or slowing at least one symptom associated with or caused by abnormal physical conditions, and Heals abnormal physical conditions. For example, treatment may be the elimination of one or more symptoms of the disorder, or the complete eradication of the disorder.

"Prevention" (i.e., the use of a compound as a precautionary measure) refers to whether it is directed to humans or animals (i.e. Prevention is concerned, in which some desired preventive effect is achieved, such as preventing disease from happening or being free from disease infection. Prevention includes preventing all symptoms of the disorder from being completely and overall from time to time, slowing down the onset of one or more symptoms of the disease, or making it less susceptible.

The prevention or treatment of a disease state or abnormal physical condition such as cancer includes slowing or reducing the incidence of cancer.

The term "mediated" as used in conjunction with IAP (also used in various physiological procedures, diseases, states, abnormal physical conditions, therapies, treatments or interventions) is intended to limit contraction, Various physiological procedures, diseases, states, abnormal physical conditions, therapies, treatments, and interventions that use the word are those who have proteins in them to play a biological role. When the term "mediated" is used in a disease, state, or abnormal physical condition, the biological effect of the protein is direct or indirect and must be and / or sufficient to cause the disease, state, or abnormal physical condition (or its cause or progression) Symptoms appear. Therefore, protein action (especially abnormal action, such as over- or under-performance) is not necessarily a proximate cause of a disease, state, or abnormal physical condition; rather, it is conceivable that such mediated diseases, states, or abnormal physical conditions include other There are multiple etiology and complex developers, in which the protein in question is only partially involved. When the term "mediating" is used for treatment, prevention, or intervention, the effect of the protein can be direct or indirect, and must and / or be sufficient to result in the treatment, prevention, or intervention. Thus, protein-mediated disease states or abnormal physical conditions include the development of resistance to any particular cancer drug or treatment.

The combination of the invention can produce a therapeutically effective effect relative to the therapeutic effect of individual compounds / agents administered individually.

The term "efficacious" includes beneficial effects such as additions, synergies, reduced side effects, reduced toxicity, increased time to disease progression, increased survival time, and sensitivity or resensitization of one agent to another ( resensitization), or increase response speed. Advantageously, the effective effect can reduce the dosage of each component to the patient, thereby reducing the toxicity of chemotherapy and producing and / or maintaining the same therapeutic effect. The term "synergistic" effect herein means that the effect produced when the agents are combined is greater than the combined effect when they are present individually. The term "additive" effect is used herein to mean that the effect produced when the agents are combined is greater than the effect when any one of them is present. "Response rate" herein, in the case of solid tumors, means the extent to which the tumor size decreases at a given time point (e.g., 12 weeks). Thus, for example, a 50% response rate means a 50% reduction in tumor size. "Clinical response" herein means a response rate of 50% or more. "Partial response" is defined herein as a response rate of less than 50%.

The term "combination" as used herein applies to two or more compounds and / or agents, and is intended to define materials in which two or more agents are associated. "Combined" and "combined" will be interpreted accordingly herein.

The association of two or more compounds and / or agents in a combination may be physical or non-physical. Examples of physically associated compounds / agents include: • a composition (e.g., a single formulation) containing two or more compounds / agents mixed together (e.g., within the same unit dose); A variety of compounds / agents are chemical / physical linkages (such as cross-linking, molecular aggregation, or bonding to a common carrier moiety); materials containing two or more compounds / agents that are chemical / physical Co-encapsulated (e.g., placed on or in a lipid vehicle, particle (e.g., micro or nano particles), or emulsion droplets); two or more compounds / agents co-encapsulated or co-encapsulated therein A pharmaceutical kit, pharmaceutical pack, or patient pack that is presented (eg, as part of a unit dose array).

Examples of non-physically associated compounds / agents include: • Materials (e.g., non-single formulations) comprising at least one of two or more compounds / agents, together with an extemporaneous association for at least one compound to Teachings to form a physical association of two or more compounds / agents; • materials (e.g., not a single formula) containing at least one of two or more compounds / agents, together with a combination therapy for two or more compounds / agents Teaching instructions; • Materials containing at least one of two or more compounds / agents, together with teachings for administration to a patient (where the other two or more compounds / agents have been (or are) being administered); The material, amount or form of at least one of the two or more compounds / agents is particularly suitable for use in combination with the other of the two or more compounds / agents.

As used herein, the term "combination therapy" is meant to include the use of a combination (as defined above) of two or more compounds / agents. Therefore, referring to "combination therapy", in the present application, "combination" and compound / agent are used in a "combination mode", which can refer to the administered compound / agent as part of the same overall treatment plan. In this regard, the dosimetry of each of the two or more compounds / agents may differ: each may be administered at the same time or at different times. Therefore, it can be seen that the compounds / agents in the combination can be administered sequentially (for example, before and after) or simultaneously, in the same pharmaceutical formula (ie, together), or in different pharmaceutical formulas (ie, individually). Those who take the same formula at the same time are a single formula, while different pharmaceutical formulas are not at the same time. The dosimetry of each of the two or more compounds / agents in the combination therapy may also differ with respect to the route of administration.

The term "pharmaceutical kit" as used herein is defined as an array of one or more unit doses of a pharmaceutical composition, together with a metering device (e.g., a measuring device) and / or a delivery device (e.g., an inhaler or syringe), and optionally All are contained in a common outer packaging. In a pharmaceutical kit containing a combination of two or more compounds / agents, the individual compounds / agents may be a single or non-single formulation. Unit doses may be contained in a blister pack. The pharmaceutical kit can optionally include teaching instructions.

As used herein, a "pharmaceutical pack" is defined as an array of one or more unit doses of a pharmaceutical composition and is optionally included in a common outer packaging. In a pharmaceutical pack containing a combination of two or more compounds / agents, the individual compounds / agents may be in a single or non-single formulation. Unit doses may be contained in a blister pack. The kit may optionally include teachings of use.

The term "n-butyl" as used herein refers to a linear alkyl group containing 4 carbon atoms.

The term "oxo" as used herein refers to a = O group.

Dotted bonds (-------) indicate single or double bonds required to complete the valence of the atoms connected by chemical bonds. It should be understood that in some cases this bond has aromatic properties. The dashed bond (-------) indicates a single or double bond, so that the ring containing X and U contains at least two double bonds.

It can be understood from the chemical formula (I) that the compound of the present invention can be represented as follows:

Wherein Q represents any of the following A, B or C:

In a specific embodiment, Q represents A. In a specific embodiment, Q represents B. In a specific embodiment, Q represents C.

In a specific embodiment, X represents CR ⁴ or N. In another embodiment, X represents CR ⁴ or NR ³ . In another embodiment, X represents N or NR ³ . In another embodiment, X represents CR ⁴ . In yet another embodiment, X represents N. In still another embodiment, X represents NR ³ .

In a specific embodiment, one of R ¹ and R ² represents hydrogen and the other represents methyl, or both R ¹ and R ² represent hydrogen. In a specific embodiment, one of R ¹ and R ² represents hydrogen and the other represents methyl. In another embodiment, R ¹ represents a methyl group and R ² represents hydrogen. In another embodiment, R ¹ represents hydrogen and R ² represents methyl. In yet another embodiment, both R ¹ and R ² represent hydrogen.

In a specific embodiment, R ³ represents hydrogen or methyl. In another embodiment, R ³ represents hydrogen or —NH ₂ . In yet another embodiment, R ³ represents methyl or —NH ₂ . In another embodiment, R ³ represents hydrogen. In yet another embodiment, R ³ represents methyl. In yet another embodiment, R ³ represents —NH ₂ .

In a specific embodiment, R ⁴ represents hydrogen or methyl. In another embodiment, R ⁴ represents hydrogen. In another embodiment, R ⁴ represents methyl.

In a specific embodiment, R ⁵ represents unsubstituted n-butyl or benzyl substituted with one or two fluorines at positions 2, 3, and / or 4 of phenyl. In a specific embodiment, R ⁵ represents unsubstituted n-butyl. In another embodiment, R ⁵ represents benzyl substituted with one or two fluorines on the phenyl group. In yet another embodiment, R ⁵ represents benzyl substituted with one or two fluorines at positions 2, 3, and / or 4 of the phenyl group. In yet another embodiment, R ⁵ represents a benzyl group substituted with a fluorine at position 2, 3, and / or 4 of the phenyl group (ie, R ⁵ represents 2-fluorobenzyl, 3-fluorobenzyl Or 4-fluorobenzyl). In yet another embodiment, R ⁵ represents a benzyl group substituted with a fluorine at position 4 of the phenyl group (ie, R ⁵ represents 4-fluorobenzyl). In another embodiment, R ⁵ represents a benzyl group substituted with two fluorines at positions 2 and 3, 3 and 4, or 2 and 4 of a phenyl group (ie, R ⁵ represents 2,3-difluoro Benzyl, 3,4-difluorobenzyl, or 2,4-difluorobenzyl). In yet another embodiment, R ⁵ represents a benzyl group substituted with two fluorines at positions 2, 4 of a phenyl group (ie, R ⁵ represents a 2,4-difluorobenzyl group).

In another embodiment, R ⁵ represents unsubstituted n-butyl, 4-fluorophenyl, or 2,4-difluorophenyl. In yet another embodiment, R ⁵ represents 4-fluorophenyl.

In a specific embodiment, R ⁶ represents methylol or -CH (OR ^x ) CH ₂ OR ^z . In a specific embodiment, R ⁶ represents methylol.

In a specific embodiment, R ⁶ represents -CH (OR ^x ) CH ₂ OR ^z . In one particular embodiment, R ^x and R ^z are hydrogen and one of the other represents a methyl group or both R ^x and R ^z are both hydrogen. In yet another embodiment, R ^x represents methyl and R ^z represents hydrogen. In another embodiment, R ^x represents hydrogen and R ^z represents methyl. In yet another embodiment, both R ^x and R ^z represent hydrogen. In yet another embodiment, R ^x represents hydrogen or methyl and R ^z represents hydrogen. In yet another embodiment, both R ^x and R ^z represent a methyl group.

In a specific embodiment, R ⁶ represents methylol, -CH (OH) CH ₂ OH, -CH (OMe) CH ₂ OH, or -CH (OH) CH ₂ OMe. In yet another embodiment, R ⁶ represents methylol, -CH (OH) CH ₂ OH, or -CH (OMe) CH ₂ OH. In yet another embodiment, R ⁶ represents hydroxymethyl.

In a specific embodiment, R ⁶ represents a pendant oxygen group (ie, = O).

Sub-Formulae

In a specific embodiment, the compound of formula (I) is: X is CR ⁴ , N, or NR ³ ; wherein, when X is CR ⁴ , U represents nitrogen and R ⁶ represents pendant oxygen; or when X is N , Then U represents carbon and R ⁶ represents methylol or -CH (OR ^x ) CH ₂ OR ^z ; or when X is NR ³ , U represents carbon and R ⁶ represents pendant oxygen; the dashed bond (---- ---) represents a single bond or a double bond, wherein at least two of the dotted bonds represent a double bond; one of R ¹ and R ² represents hydrogen and the other represents a methyl group or both R ¹ and R ² represent Hydrogen; R ³ represents hydrogen, methyl or -NH ₂ ; R ⁴ represents hydrogen or methyl; R ⁵ represents unsubstituted n-butyl or one or two at positions 2, 3 and / or 4 of phenyl the fluoro-substituted benzyl; and R ^x represent, and R ^z represents hydrogen and the other one of those indicated both methyl or R ^x and R ^z are both hydrogen.

In another specific embodiment, the compound of formula (I) is: X is CR ⁴ , N or NR ³ ; wherein when X is CR ⁴ , U represents nitrogen and R ⁶ represents a pendant oxygen group; or when X is CR N, then U represents carbon and R ⁶ represents methylol or -CH (OR ^x ) CH ₂ OR ^z ; or when X is NR ³ , U represents carbon and R ⁶ represents side oxygen; dashed bond (--- ----) means a single bond or a double bond where at least two of the dotted bonds represent a double bond; one of R ¹ and R ² represents hydrogen and the other represents a methyl group or both R ¹ and R ² represent Hydrogen; R ³ represents hydrogen, methyl or -NH ₂ ; R ⁴ represents hydrogen or methyl; R ⁵ represents unsubstituted n-butyl, 4-fluorobenzyl or 2,4-fluorobenzyl; R ^x represents hydrogen or methyl; and R ^z represents hydrogen.

In a specific embodiment, the compound of formula (I) is a compound of formula (Ia): (Ia) or a tautomer thereof, a stereochemical isomer thereof, a pharmaceutically acceptable salt thereof, or a solvate thereof; wherein R ¹ , R ² , R ^4, and R ^{5 are} any of the specific examples One is defined.

In a specific embodiment of the compound of formula (Ia), one of R ¹ and R ² represents hydrogen and the other represents methyl or R ¹ and R ² both represent hydrogen. In another specific embodiment of the compound of formula (Ia), R ¹ represents hydrogen and R ² represents methyl or R ¹ and R ² both represent hydrogen.

In yet another embodiment of the compound of formula (Ia), R ¹ represents methyl and R ² represents hydrogen. In another embodiment of the compound of formula (Ia), R ¹ represents hydrogen and R ² represents methyl.

In a specific embodiment of the compound of formula (Ia), R ⁴ represents hydrogen or methyl.

In a specific embodiment of the compound of formula (Ia), R ⁵ represents unsubstituted n-butyl or benzyl substituted with one or two fluorines at positions 2, 3 and / or 4 of phenyl. In a specific embodiment of the compound of formula (Ia), R ⁵ represents unsubstituted n-butyl. In another embodiment of the compound of formula (Ia), R ⁵ represents benzyl substituted with one or two fluorines on the phenyl group. In yet another embodiment of the compound of formula (Ia), R ⁵ represents benzyl substituted with one or two fluorines at positions 2, 3, and / or 4 of the phenyl group. In yet another embodiment of the compound of formula (Ia), R ⁵ represents benzyl substituted with a fluorine at position 4 of the phenyl group (ie, R ⁵ represents 4-fluorobenzyl). In yet another embodiment of the compound of formula (Ia), R ⁵ represents a benzyl group substituted with two fluorines at positions 2, 4 of the phenyl group (ie, R ⁵ represents a 2,4-difluorobenzyl group) ).

In a specific embodiment, the compound of formula (I) is a compound of formula (Ib): (Ib) or a tautomer thereof, a stereochemical isomer thereof, a pharmaceutically acceptable salt thereof, or a solvate thereof; wherein R ¹ , R ² , R ⁵ , R ⁶ , R ^x and R ^z As described in any of these specific embodiments. In a specific embodiment, R ⁶ represents methylol or -CH (OR ^x ) CH ₂ OR ^z .

In a specific embodiment of the compound of formula (Ib), R ¹ represents methyl and R ² represents hydrogen or both R ¹ and R ² represent hydrogen.

In another embodiment of the compound of formula (Ib), both R ¹ and R ² represent hydrogen.

In a specific embodiment of the compound of formula (Ib), R ⁵ represents unsubstituted n-butyl or benzyl substituted with one or two fluorines at positions 2, 3, and / or 4 of phenyl. In a specific embodiment of the compound of formula (Ib), R ⁵ represents unsubstituted n-butyl. In another embodiment of the compound of formula (Ib), R ⁵ represents benzyl substituted with one or two fluorines on the phenyl group. In yet another embodiment of the compound of formula (Ib), R ⁵ represents benzyl substituted with one or two fluorines at positions 2, 3, and / or 4 of the phenyl group. In yet another embodiment of the compound of formula (Ib), R ⁵ represents a benzyl group substituted with a fluorine at position 2, 3, and / or 4 of the phenyl group (ie, R ⁵ represents 2-fluorobenzyl , 3-fluorobenzyl or 4-fluorobenzyl). In yet another embodiment of the compound of formula (Ib), R ⁵ represents a benzyl group substituted with two fluorines at positions 2, 4 of a phenyl group (ie, R ⁵ represents a 2,4-difluorobenzyl group) ). In still another specific embodiment of the compound of formula (Ib), R ⁵ represents a benzyl group substituted with a fluorine at position 4 of the phenyl group (ie, R ⁵ represents 4-fluorobenzyl).

In a specific embodiment of the compound of formula (Ib), R ⁶ represents methylol, -CH (OH) CH ₂ OH, -CH (OMe) CH ₂ OH, or -CH (OH) CH ₂ OMe.

In another specific embodiment of the compound of formula (Ib), R ⁶ represents methylol, -CH (OH) CH ₂ OH, or -CH (OMe) CH ₂ OH.

In yet another embodiment of the compound of formula (Ib), R ⁶ represents hydroxymethyl.

In a specific embodiment, the compound of formula (I) is a compound of formula (Ic): (Ic) or a tautomer thereof, a stereochemical isomer thereof, a pharmaceutically acceptable salt thereof, or a solvate thereof; wherein R ¹ , R ² , R ⁴ and R ^{5 are} any of the specific examples One is defined.

In a specific embodiment of the compound of formula (Ic), one of R ¹ and R ² represents hydrogen and the other represents methyl or R ¹ and R ² both represent hydrogen. In another embodiment of the compound of formula (Ic), R ¹ represents a methyl group and R ² represents hydrogen or both R ¹ and R ² represent hydrogen.

In a specific embodiment of the compound of formula (Ic), R ³ represents hydrogen or methyl. In another embodiment of the compound of formula (Ic), R ³ represents hydrogen or -NH ₂ . In yet another embodiment of the compound of formula (Ic), R ³ represents methyl or -NH ₂ . In another embodiment of the compound of formula (Ic), R ³ represents hydrogen. In yet another embodiment of the compound of formula (Ic), R ³ represents methyl. In yet another embodiment of the compound of formula (Ic), R ³ represents -NH ₂ .

In a specific embodiment of the compound of formula (Ic), R ⁵ represents unsubstituted n-butyl or benzyl substituted with one or two fluorines at positions 2, 3, and / or 4 of phenyl. In a specific embodiment of the compound of formula (Ic), R ⁵ represents unsubstituted n-butyl. In another embodiment of the compound of formula (Ic), R ⁵ represents benzyl substituted with one or two fluorines on the phenyl group. In yet another embodiment of the compound of formula (Ic), R ⁵ represents benzyl substituted with one or two fluorines at positions 2, 3, and / or 4 of the phenyl group. In another specific embodiment of the compound of formula (Ic), R ⁵ represents a benzyl group substituted with a fluorine at position 2 or 4 of the phenyl group (ie, R ⁵ represents 2-fluorobenzyl or 4-fluoro Benzyl). In yet another embodiment of the compound of formula (Ic), R ⁵ represents benzyl substituted with a fluorine at position 2 of the phenyl group (ie, R ⁵ represents 4-fluorobenzyl). In another embodiment of the compound of formula (Ic), R ⁵ represents a benzyl group substituted with two fluorines at positions 2, 4 of the phenyl group (ie, R ⁵ represents a 2,4-difluorobenzyl group) ).

In a specific embodiment, the compound of formula (I) is a compound of formula (Id): (Id) or a tautomer thereof, a stereochemical isomer thereof, a pharmaceutically acceptable salt thereof, or a solvate thereof; wherein R ^{5 is} as defined in any one of the specific embodiments.

In a specific embodiment of the compound of formula (Id), R ⁵ represents unsubstituted n-butyl or benzyl substituted with one or two fluorines at positions 2, 3 and / or 4 of phenyl. In a specific embodiment of a compound of formula (Id), R ⁵ represents unsubstituted n-butyl. In another embodiment of the compound of formula (Id), R ⁵ represents benzyl substituted with one or two fluorines on the phenyl group. In another embodiment of the compound of formula (Id), R ⁵ represents benzyl substituted with one or two fluorines at positions 2, 3, and / or 4 of the phenyl group. In another specific embodiment of the compound of formula (Id), R ⁵ represents a benzyl group substituted with a fluorine at position 2, 3, and / or 4 of the phenyl group (ie, R ⁵ represents 2-fluorobenzyl , 3-fluorobenzyl or 4-fluorobenzyl). In another specific embodiment of the compound of formula (Id), R ⁵ represents a benzyl group substituted with two fluorines at positions 2, 4 of a phenyl group (ie, R ⁵ represents a 2,4-difluorobenzyl group) ). In yet another embodiment of the compound of formula (Id), R ⁵ represents a benzyl group substituted with a fluorine at position 4 of the phenyl group (ie, R ⁵ represents 4-fluorobenzyl).

In a specific embodiment, the compound of formula (I) is a compound of formula (I), (Ia), (Ib) or (Ic), wherein R ¹ represents a methyl group and R ² represents hydrogen.

In a specific embodiment, the compound of formula (I) is a compound of formula (I), (Ia), (Ib) or (Ic), wherein R ¹ and R ² both represent hydrogen.

In a specific embodiment, the compound of formula (I) is a compound of formula (I), (Ia), (Ib) or (Ic), wherein R ¹ represents hydrogen and R ² represents methyl.

In a specific embodiment, the compound of formula (I) is a compound of formula (I), (Ia), (Ib), (Ic) or (Id), wherein R ⁵ represents unsubstituted n-butyl.

In a specific embodiment, the compound of formula (I) is a compound of formula (I), (Ia), (Ib), (Ic) or (Id), wherein R ⁵ represents the position 2, 3 and / or of the phenyl group 4 has one or two fluorine-substituted benzyl groups.

In a specific embodiment, the compound of formula (I) is a compound of formula (I), (Ia), (Ib), (Ic) or (Id), wherein R ⁵ represents one or two fluorines at the position of the phenyl group. Substituted benzyl.

In a specific embodiment, the compound of formula (I) is a compound of formula (I), (Ia), (Ib), (Ic) or (Id), wherein R ⁵ represents benzene having two fluorine-substituted benzenes on the phenyl group. Methyl, such as 2,3 disubstituted, 2,4 disubstituted, 2,5 disubstituted, 3,5 disubstituted, 2,6 disubstituted, or 3,4 disubstituted.

In a specific embodiment, the compound of formula (I) is a compound of formula (I), (Ia), (Ib), (Ic) or (Id), wherein R ⁵ represents the position 2, 3 and / or of the phenyl group 4 has one or two fluorine-substituted benzyl groups.

In a specific embodiment, the compound of formula (I) is a compound of formula (I), (Ia), (Ib), (Ic) or (Id), wherein R ⁵ represents the position 2, 3 or 4 of the phenyl group. There is one benzyl substituted by fluorine (ie, R ⁵ represents 2-fluorobenzyl, 3-fluorobenzyl or 4-fluorobenzyl).

In a specific embodiment, the compound of formula (I) is a compound of formula (I), (Ia), (Ib), (Ic) or (Id), wherein R ⁵ represents the positions 2 and 3, 3 and 4, or 2 and 4 benzyl substituted with two fluorines (ie, R ⁵ represents 2,3-difluorobenzyl, 3,4-difluorobenzyl, or 2,4-difluorobenzyl ).

In a specific embodiment, the compound of the formula (I) is a compound of the formula (I), (Ia), (Ib), (Ic) or (Id), wherein R ⁵ represents two groups at positions 2 and 4 of the phenyl group. Fluoro-substituted benzyl (ie, R ⁵ represents 2,4-difluorobenzyl).

In a specific embodiment, the compound of formula (I) is a compound of formula (I), (Ia), (Ib), (Ic) or (Id), wherein R ⁵ represents 2,4-difluorobenzyl or 4 -Fluorobenzyl.

In a specific embodiment, the present invention provides a compound of formula (I), which includes a free base or a tautomer of the compound of Examples 1 to 37, a stereochemical isomer thereof, a pharmaceutically acceptable salt thereof, or Its solvate.

In a specific embodiment, the present invention provides a compound of formula (I), which is a free base or a tautomer of the compound of Examples 1 to 37, a stereochemical isomer thereof, a pharmaceutically acceptable salt thereof, or Its solvate.

In a specific embodiment, the present invention provides a compound of formula (I), which includes the compounds of Examples 1 to 37 or a tautomer thereof, a stereochemical isomer thereof, or a solvate thereof.

In yet another embodiment, the compound is a free base or a tautomer thereof, a stereochemical isomer thereof, a pharmaceutically acceptable salt thereof, or a solvate thereof selected from Examples 1 to 34.

In yet another embodiment, the present invention provides a compound of formula (I), which includes a compound selected from the following:

1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridine- 1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} piperazin-1-yl] ethyl- 1-keto (1- {6-[(4-Fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} piperazin-1-yl] ethan-1-one);

6-[(4-fluorophenyl) methyl] -3,3,4-trimethyl-1- {2-[(2R, 5R) -5-methyl-2-{[((3R) -3 -Methylmorpholin-4-yl] methyl} piperazin-1-yl] ethenyl} -1H, 2H, 3H, 4H, 5H-pyrrolo [3,2-b] pyridin-5-one ( 6-[(4-Fluorophenyl) methyl] -3,3,4-trimethyl-1- {2-[(2R, 5R) -5-methyl-2-{[(3R) -3-methylmorpholin-4-yl ] methyl} piperazin-1-yl] acetyl} -1H, 2H, 3H, 4H, 5H-pyrrolo [3,2-b] pyridin-5-one);

6-[(2,4-difluorophenyl) methyl] -3,3-dimethyl-1- {2-[(2R, 5R) -5-methyl-2-{[((3R)- 3-methylmorpholin-4-yl] methyl} piperazin-1-yl] ethenyl} -1H, 2H, 3H, 4H, 5H-pyrrolo [3,2-b] pyridin-5-one (6-[(2,4-Difluorophenyl) methyl] -3,3-dimethyl-1- {2-[(2R, 5R) -5-methyl-2-{[(3R) -3-methylmorpholin-4- yl] methyl} piperazin-1-yl] acetyl} -1H, 2H, 3H, 4H, 5H-pyrrolo [3,2-b] pyridin-5-one);

6-[(2,4-difluorophenyl) methyl] -3,3,4-trimethyl-1- {2-[(2R, 5R) -5-methyl-2-{[((3R ) -3-methylmorpholin-4-yl] methyl} piperazin-1-yl] ethenyl} -1H, 2H, 3H, 4H, 5H-pyrrolo [3,2-b] pyridine-5 -Keto (6-[(2,4-Difluorophenyl) methyl] -3,3,4-trimethyl-1- {2-[(2R, 5R) -5-methyl-2-{[(3R) -3- methylmorpholin-4-yl] methyl} piperazin-1-yl] acetyl} -1H, 2H, 3H, 4H, 5H-pyrrolo [3,2-b] pyridin-5-one);

1- [5-((R or S) -1,2-dihydroxyethyl) -6-[(4-fluorophenyl) methyl] -3,3-dimethyl-1H, 2H, 3H- Pyrrolo [3,2-b] pyridin-1-yl] -2-[(2R, 5R) -2-{[((3R, 5R) -3,5-dimethylmorpholin-4-yl] methyl } -5-methylpiperazin-1-yl] ethyl-1-one (1- [5-((R or S) -1,2-Dihydroxyethyl) -6-[(4-fluorophenyl) methyl]- 3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl] -2-[(2R, 5R) -2-{[(3R, 5R) -3,5- dimethylmorpholin-4-yl] methyl} -5-methylpiperazin-1-yl] ethan-1-one);

6-[(2,4-difluorophenyl) methyl] -1- {2-[(2R, 5R) -2-{[((2S, 5R) -2,5-dimethylmorpholine-4 -Yl] methyl} -5-methylpiperazin-1-yl] ethenyl} -3,3-dimethyl-1H, 2H, 3H, 5H, 6H-pyrrolo [2,3-c] Pyridine-5-one (6-[(2,4-Difluorophenyl) methyl] -1- {2-[(2R, 5R) -2-{[(2S, 5R) -2,5-dimethylmorpholin-4-yl ] methyl} -5-methylpiperazin-1-yl] acetyl} -3,3-dimethyl-1H, 2H, 3H, 5H, 6H-pyrrolo [2,3-c] pyridin-5-one);

4-amino-6-[(4-fluorophenyl) methyl] -3,3-dimethyl-1- {2-[(2R, 5R) -5-methyl-2-{[(3R ) -3-methylmorpholin-4-yl] methyl} piperazin-1-yl] ethenyl} -1H, 2H, 3H, 4H, 5H-pyrrolo [3,2-b] pyridine-5 -Keto (4-Amino-6-[(4-fluorophenyl) methyl] -3,3-dimethyl-1- {2-[(2R, 5R) -5-methyl-2-{[(3R) -3- methylmorpholin-4-yl] methyl} piperazin-1-yl] acetyl} -1H, 2H, 3H, 4H, 5H-pyrrolo [3,2-b] pyridin-5-one);

1- {6-[(4-fluorophenyl) methyl] -5-((R or S) -2-hydroxy-1-methoxyethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl ] Methyl} piperazin-1-yl] ethyl-1-one (1- {6-[(4-Fluorophenyl) methyl] -5-((R or S) -2-hydroxy-1-methoxyethyl) -3 , 3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[(3R) -3-methylmorpholin -4-yl] methyl} piperazin-1-yl] ethan-1-one);

4-amino-6-butyl-1- {2-[(2R, 5R) -2-{[((3R, 5R) -3,5-dimethylmorpholin-4-yl] methyl}- 5-methylpiperazin-1-yl] ethenyl} -3,3-dimethyl-1H, 2H, 3H, 4H, 5H-pyrrolo [3,2-b] pyridin-5-one (4 -Amino-6-butyl-1- {2-[(2R, 5R) -2-{[(3R, 5R) -3,5-dimethylmorpholin-4-yl] methyl} -5-methylpiperazin-1-yl] acetyl} -3,3-dimethyl-1H, 2H, 3H, 4H, 5H-pyrrolo [3,2-b] pyridin-5-one);

6-[(2,4-difluorophenyl) methyl] -3,3,4-trimethyl-1- {2-[(2R, 5R) -5-methyl-2-{[((3R ) -3-methylmorpholin-4-yl] methyl} piperazin-1-yl] ethenyl} -1H, 2H, 3H, 5H, 6H-pyrrolo [2,3-c] pyridine-5 -Keto (6-[(2,4-Difluorophenyl) methyl] -3,3,4-trimethyl-1- {2-[(2R, 5R) -5-methyl-2-{[(3R) -3- methylmorpholin-4-yl] methyl} piperazin-1-yl] acetyl} -1H, 2H, 3H, 5H, 6H-pyrrolo [2,3-c] pyridin-5-one);

6-butyl-1- {2-[(2R, 5R) -2-{[((3R, 5R) -3,5-dimethylmorpholin-4-yl] methyl} -5-methylpiperazine Azin-1-yl] ethenyl} -3,3-dimethyl-1H, 2H, 3H, 4H, 5H-pyrrolo [3,2-b] pyridin-5-one (6-Butyl-1- {2-[(2R, 5R) -2-{[((3R, 5R) -3,5-dimethylmorpholin-4-yl] methyl} -5-methylpiperazin-1-yl] acetyl} -3,3-dimethyl- 1H, 2H, 3H, 4H, 5H-pyrrolo [3,2-b] pyridin-5-one); and

6-butyl-1- {2-[(2R, 5R) -2-{[((2S, 5R) -2,5-dimethylmorpholin-4-yl] methyl} -5-methylpiperazine Azin-1-yl] ethenyl} -3,3-dimethyl-1H, 2H, 3H, 4H, 5H-pyrrolo [3,2-b] pyridin-5-one (6-Butyl-1- {2-[(2R, 5R) -2-{[((2S, 5R) -2,5-dimethylmorpholin-4-yl] methyl} -5-methylpiperazin-1-yl] acetyl} -3,3-dimethyl- 1H, 2H, 3H, 4H, 5H-pyrrolo [3,2-b] pyridin-5-one);

Or a tautomer thereof, a stereochemical isomer thereof, a pharmaceutically acceptable salt thereof, or a solvate thereof.

In another embodiment, the present invention provides a compound selected from the group consisting of:

1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridine- 1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} piperazin-1-yl] ethyl- 1-ketodihydrochloride (1- {6-[(4-Fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin -1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} piperazin-1-yl] ethan-1-one dihydrochloride) (E2);

6-[(4-fluorophenyl) methyl] -3,3,4-trimethyl-1- {2-[(2R, 5R) -5-methyl-2-{[((3R) -3 -Methylmorpholin-4-yl] methyl} piperazin-1-yl] ethenyl} -1H, 2H, 3H, 4H, 5H-pyrrolo [3,2-b] pyridin-5-one di Hydrochloride (E6);

6-[(2,4-difluorophenyl) methyl] -3,3-dimethyl-1- {2-[(2R, 5R) -5-methyl-2-{[((3R)- 3-methylmorpholin-4-yl] methyl} piperazin-1-yl] ethenyl} -1H, 2H, 3H, 4H, 5H-pyrrolo [3,2-b] pyridin-5-one Dihydrochloride (E8);

6-[(2,4-difluorophenyl) methyl] -3,3,4-trimethyl-1- {2-[(2R, 5R) -5-methyl-2-{[((3R ) -3-methylmorpholin-4-yl] methyl} piperazin-1-yl] ethenyl} -1H, 2H, 3H, 4H, 5H-pyrrolo [3,2-b] pyridine-5 -Ketodihydrochloride (E19);

1- [5-((R or S) -1,2-dihydroxyethyl) -6-[(4-fluorophenyl) methyl] -3,3-dimethyl-1H, 2H, 3H- Pyrrolo [3,2-b] pyridin-1-yl] -2-[(2R, 5R) -2-{[((3R, 5R) -3,5-dimethylmorpholin-4-yl] methyl } -5-methylpiperazin-1-yl] ethyl-1-one dihydrochloride (E21);

6-[(2,4-difluorophenyl) methyl] -1- {2-[(2R, 5R) -2-{[((2S, 5R) -2,5-dimethylmorpholine-4 -Yl] methyl} -5-methylpiperazin-1-yl] ethenyl} -3,3-dimethyl-1H, 2H, 3H, 5H, 6H-pyrrolo [2,3-c] Pyridine-5-one dihydrochloride (E22);

4-amino-6-[(4-fluorophenyl) methyl] -3,3-dimethyl-1- {2-[(2R, 5R) -5-methyl-2-{[(3R ) -3-methylmorpholin-4-yl] methyl} piperazin-1-yl] ethenyl} -1H, 2H, 3H, 4H, 5H-pyrrolo [3,2-b] pyridine-5 -Ketodihydrochloride (E24);

1- {6-[(4-fluorophenyl) methyl] -5-((R or S) -2-hydroxy-1-methoxyethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl ] Methyl} piperazin-1-yl] ethyl-1-one trihydrochloride (E27);

4-amino-6-butyl-1- {2-[(2R, 5R) -2-{[((3R, 5R) -3,5-dimethylmorpholin-4-yl] methyl}- 5-methylpiperazin-1-yl] ethenyl} -3,3-dimethyl-1H, 2H, 3H, 4H, 5H-pyrrolo [3,2-b] pyridin-5-onedihydro Chlorate (E30);

6-[(2,4-difluorophenyl) methyl] -3,3,4-trimethyl-1- {2-[(2R, 5R) -5-methyl-2-{[((3R ) -3-methylmorpholin-4-yl] methyl} piperazin-1-yl] ethenyl} -1H, 2H, 3H, 5H, 6H-pyrrolo [2,3-c] pyridine-5 -Ketodihydrochloride (E31);

6-butyl-1- {2-[(2R, 5R) -2-{[((3R, 5R) -3,5-dimethylmorpholin-4-yl] methyl} -5-methylpiperazine Azin-1-yl] ethenyl} -3,3-dimethyl-1H, 2H, 3H, 4H, 5H-pyrrolo [3,2-b] pyridin-5-one dihydrochloride (E32 )and

6-butyl-1- {2-[(2R, 5R) -2-{[((2S, 5R) -2,5-dimethylmorpholin-4-yl] methyl} -5-methylpiperazine Azin-1-yl] ethenyl} -3,3-dimethyl-1H, 2H, 3H, 4H, 5H-pyrrolo [3,2-b] pyridin-5-one dihydrochloride (E37 )

Or a tautomer thereof, a stereochemical isomer thereof, or a solvate thereof.

In another specific embodiment, the compound is a free base selected from Examples 2, 6, 19, 21, 22, 24, 27, 30, 31, and 32, or a tautomer, a stereochemical isomer thereof, Its pharmaceutically acceptable salt, or its solvate.

In yet another specific embodiment, the present invention provides a compound of formula (I), which comprises 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-di Methyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[(3R) -3-form Morpholin-4-yl] methyl} piperazin-1-yl] ethyl-1-one (1- {6-[(4-Fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl- 1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[(3R) -3-methylmorpholin-4-yl] methyl} piperazin-1-yl] ethan-1-one) or a tautomer thereof, a stereochemical isomer thereof, a pharmaceutically acceptable salt thereof, or a solvate thereof.

In yet another specific embodiment, the present invention provides a compound of formula (I), which comprises 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-di Methyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[(3R) -3-form Morpholin-4-yl] methyl} piperazin-1-yl] ethan-1-one hydrochloride or a tautomer thereof, a stereochemical isomer thereof, or a solvate thereof.

In yet another specific embodiment, the present invention provides a compound of formula (I), which comprises 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-di Methyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[(3R) -3-form Morpholin-4-yl] methyl} piperazin-1-yl] ethan-1-one dihydrochloride (E2).

In still another specific embodiment, the present invention provides 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H -Pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[(3R) -3-methylmorpholin-4-yl] Methyl} piperazin-1-yl] ethyl-1-one lactate (1- {6-[(4-Fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H- pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[(3R) -3-methylmorpholin-4-yl] methyl} piperazin-1- yl] ethan-1-one lactate salt) or a tautomer thereof, a stereochemical isomer thereof, or a solvate thereof.

In still another specific embodiment, the present invention provides 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H -Pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[(3R) -3-methylmorpholin-4-yl] Methyl} piperazin-1-yl] ethyl-1-one L-(+)-lactate (1- {6-[(4-Fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl- 1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[(3R) -3-methylmorpholin-4-yl] methyl} piperazin-1-yl] ethan-1-one L-(+)-lactate salt) or a tautomer thereof, a stereochemical isomer thereof, or a solvate thereof.

In still another specific embodiment, the compound is selected from Examples 38-42.

In still another specific embodiment, the present invention provides 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H -Pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[(3R) -3-methylmorpholin-4-yl] Methyl} piperazin-1-yl] ethyl-1-one L-(+)-lactate (type A) (E39).

In still another specific embodiment, the present invention provides 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H -Pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[(3R) -3-methylmorpholin-4-yl] Methyl} piperazin-1-yl] ethan-1-one L-(+)-lactate (type B) (E40).

In still another specific embodiment, the present invention provides 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H -Pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[(3R) -3-methylmorpholin-4-yl] Methyl} piperazin-1-yl] ethan-1-one L-(+)-lactate (type C) (E43).

In still another specific embodiment, the present invention provides 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H -Pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[(3R) -3-methylmorpholin-4-yl] Methyl} piperazin-1-yl] ethyl-1-one sulfate (1- {6-[(4-Fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H- pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[(3R) -3-methylmorpholin-4-yl] methyl} piperazin-1- yl] ethan-1-one sulfate salt) or a tautomer thereof, a stereochemical isomer thereof, or a solvate thereof.

In still another specific embodiment, the present invention provides 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H -Pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[(3R) -3-methylmorpholin-4-yl] Methyl} piperazin-1-yl] ethan-1-one sulfate (Form F) (E41).

In still another specific embodiment, the present invention provides 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H -Pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[(3R) -3-methylmorpholin-4-yl] Methyl} piperazin-1-yl] ethan-1-one methylsulfonate (1- {6-[(4-Fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H , 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[(3R) -3-methylmorpholin-4-yl] methyl} piperazin -1-yl] ethan-1-one mesylate salt) or a tautomer thereof, a stereochemical isomer thereof, or a solvate thereof.

In still another specific embodiment, the present invention provides 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H -Pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[(3R) -3-methylmorpholin-4-yl] Methyl} piperazin-1-yl] ethyl-1-one mesylate (type B) (E42).

In another embodiment, the selected compound is a compound other than Example 35, or a tautomer, a stereochemical isomer thereof, a pharmaceutically acceptable salt thereof, or a solvate thereof.

In another specific embodiment, the selected compound is other than Example 2, or a tautomer, a stereochemical isomer thereof, a pharmaceutically acceptable salt thereof, or a solvate thereof.

For the avoidance of doubt, it should be understood that the general and specific preferred examples, embodiments, and examples of a substituent may be combined with one or more substituents, particularly all other substituents as defined herein Combined with specific preferred examples, embodiments, and examples, all of which are covered by this application.

Salts, solvates, tautomers, isomers, N-oxides, esters, prodrugs, and isotopes

The compounds of formula (I) and their subgroups also include their ionic forms, salts, solvates, isomers (including geometric isomers and stereochemical isomers), tautomers, N-oxidation Compounds, esters, prodrugs, isotopes, and protected forms thereof, such as the following; preferably their salts or tautomers or isomers or N-oxides or solvates; more preferably their salts or tautomers Structure or N-oxide or solvate; even more preferably its salt or tautomer or solvate.

salt

Various compounds of formula (I) may exist in the form of salts, for example, acid addition salts, or salts of organic and inorganic bases, such as carboxylates, sulfonates and phosphates, in specific examples. All such salts are within the scope of this invention, and references to compounds of formula (I) may include the salt forms of these compounds.

The salt of the present invention can be synthesized from the parent compound containing basic or acidic groups by conventional chemical methods, such as Pharmaceutical Salts: Properties, Selection, and Use, P. Heinrich Stahl (Editor), CamilleG. Wermuth ( Editor), ISBN: 3-90639-026-8, Hardcover, 388 pages, August 2002. In general, such salts can be obtained by reacting the free acid or base form of a compound with an appropriate base or acid in water or an organic solution or a mixture thereof; in general, a non-aqueous solute is used , Such as ether, ethyl acetate, ethanol, isopropanol, or propionitrile.

It can form acid addition salts (mono or double salts) with a wide range of acids (including inorganic and organic acids). Examples of acid addition salts include single or double salts with acids, and the acid is selected from the group consisting of acetic acid, 2,2-dichloroacetic acid, adipic, alginic, ascorbic (such as , L-ascorbic acid), L-aspartic, benzenesulfonic, benzoic, 4-acetamidobenzoic, butanoic , (+) Camphoric acid ((+) camphoric), camphor-sulfonic, (+)-(1S) -camphor-10-sulfonic acid, capric, caproic, caprylic acid (caprylic), cinnamic, citric, cyclamic, dodecylsulfuric, ethane-1,2-disulfonic , Ethanesulfonic, 2-hydroxyethanesulfonic, formic, fumaric, galactaric, gentisic, glucoheptose Glucoheptonic, D-gluconic, glucuronic (e.g., D-glucuronic acid), glutamic (e.g., L-glutamic acid), α-oxoglutaric, glycolic, Hippuric, hydrohalic acid (e.g., hydrobromic acid, hydrochloric acid, hydroiodic acid), isethionic, lactic acid (e.g., (+)-L-lactic acid , (±) -DL-lactic acid), lactobionic, maleic, malic, (-)-L-malic acid, malonic, (±) -DL -Almond acid ((±) -DL-mandelic), methanesulfonic, naphthalene-2-sulfonic, naphthalene-1,5- disulfonic), 1-hydroxy-2-naphthoic, nicotinic, nitric acid, oleic, orotic, oxalic, palmitic acid (palmitic), pamoic, phosphoric, propionic, pyricic, L-pyroglutamic, salicylic, 4-amino -Salicylic acid (4-amino-salicylic), sebacic, stearic, succinic, sulfuric, tannic, (+)-L-tartaric acid ((+)-L-tartaric), thiocyanic, p- toluencsulfonic, undecylenic, and valeric acid ), And acylated amino acids and cation exchange resins.

A special salt group is composed of the following acids: acetic acid, hydrochloric acid, hydroiodic acid, phosphoric acid, nitric acid, sulfuric acid, citric acid, lactic acid, succinic acid, maleic acid, malic acid, Isethionate, fumaric acid, benzenesulfonic acid, toluenesulfonic acid, methanesulfonic acid (also known as mesylate), ethanesulfonic acid, naphthalenesulfonic acid, valeric acid, acetic acid, propionic acid, Butyric acid, malonic acid, glucuronic acid and lactobionic acid. A special salt subgroup is composed of salts of the following acids: hydrochloric acid, lactic acid (such as (+)-L-lactic acid, (-)-D-lactic acid, or (±) -DL- Lactic acid), sulfuric acid, and methanesulfonic acid (methanesulfonic acid). A more specific subgroup of salts is composed of salts of the following acids: lactic acid (e.g., (+)-L-lactic acid, (-)-D-lactic acid or (±) -DL-lactic acid) and sulfuric acid. A special salt is the hydrochloride. A more specific salt is lactate (such as the compounds of Examples 39, 40, and 43). A more specific salt is sulfate (e.g., the compound of Example 41). A more specific salt is a mesylate (e.g., the compound of Example 42). A more specific salt is lactate (such as the compounds of Examples 39, 40, and 43, especially the compound of Example 43), such as L-(+)-lactate.

If the compound is anionic, or has a can when a functional group of the anionic (e.g., -COOH may be -COO ^-), and can be adapted to produce a cation of an organic or inorganic salts with bases. Examples of suitable inorganic cations include, but are not limited to, alkali metal cations such as Li ⁺ , Na ⁺ and K ⁺ , alkaline earth metal cations such as Ca ²⁺ and Mg ²⁺ , and other cations such as Al ³⁺ or Zn ⁺ . Examples of suitable organic cations include, but are not limited to: ammonium ions (ie, NH ⁴⁺ ) and substituted ammonium ions (eg, NH ₃ R ⁺ , NH ₂ R ₂ ⁺ , NHR ₃ ⁺ , NR ₄ ⁺ ). Examples of some suitable substituted ammonium ions are derived from: methylamine, ethylamine, diethylamine, propylamine, dicyclohexylamine, triethylamine, butylamine, ethylenediamine , Ethanolamine, diethanolamine, piperazine, benzylamine, phenylbenzylamine, choline, meglumine and tromethamine, and amino acids such as lysine and Arginine. An example of a common quaternary ammonium ion is N (CH ₃ ) ₄ ⁺ .

When the compound of formula (I) contains a monoamine functional group, a quaternary ammonium salt can be formed, for example, by reacting with an alkylating agent to form a quaternary ammonium salt according to methods known to those skilled in the art. Such quaternary ammonium salt compounds are also included in the scope of Chemical Formula (I).

The compounds of the present invention may exist as mono-, bis- or tri-salts, especially mono- or bis-salts, depending on the pKa value of the acid which forms the salt.

The salt form of the compound of the present invention is a general pharmaceutically acceptable salt, and examples of the pharmaceutically acceptable salt are Berge et al., 1977, "Pharmaceutically Acceptable Salts," J. Pharm. Sci., Vol. 66 , pp.1-19. However, non-pharmaceutically acceptable salts can also be prepared as an intermediate form compound for subsequent conversion to pharmaceutically acceptable salts. Such non-pharmaceutically acceptable salts are also useful, for example, when purifying or isolating compounds of the invention, and are part of the invention.

In a specific embodiment of the present invention, a pharmaceutical composition is provided, which includes a solution (eg, an aqueous solution) containing a compound of formula (I) and a subgroup thereof. In the example described herein, the salt concentration is not In excess of 10 mg / ml, it is generally greater than 15 mg / ml, and preferably greater than 20 mg / ml.

N-oxide

Compounds of formula (I) containing amine functional groups can also form N-oxides. The amine functional group-containing compound of formula (I) referred to herein also includes N-oxides.

When a compound contains multiple amine functional groups, one or more nitrogen atoms can be oxidized to form an N-oxide. Specific examples of N-oxides are tertiary amines or N-oxides of nitrogen atoms of nitrogen-containing heterocycles.

N-oxides can be formed by treating the corresponding amine with an oxidant such as hydrogen peroxide or per-acid (such as peroxycarboxylic acid), such as Advanced Organic Chemistry, by Jerry March, 4 ^th Edition, Wiley Interscience, pages. More preferably, the N-oxide can be produced by a process described in LW Deady (Syn. Comm. 1977, 7, 509-514), in which an amine compound and m-chloroperoxybenzoic acid (MCPBA) are used in The reaction is carried out in an inert solvent such as dichloromethane.

Geometric isomers or tautomers

Compounds of formula (I) may also exist in a variety of different geometric isomers and tautomers, and the compounds of formula (I) referred to include all such forms. For the avoidance of doubt, compounds may exist in one of a variety of geometric isomers or tautomer forms, and only one of them is specifically described or shown, however, other forms are also included within the scope of formula (I).

For example, in the compound of formula (I), when X represents NH and U represents carbon, the benzene ring of the compound can exist in tautomeric forms as shown below. For the sake of brevity, the general formula (I) shows a form 1, but this general formula is considered to include two tautomeric forms (1 and 2).

1 2

Other examples of tautomeric forms include, for example, keto-, enol-, and enolate-forms, such as the following paired tautomers: keto / enol (below (Shown), imine / enamine, imine / enamine, amidine / enediamine, nitroso / oxime, thioketone / enethiol ( thioketone / enethiol) and nitro / aci-nitro.

Stereoisomers

Unless otherwise mentioned or stated, the chemical designation of a compound means a mixture of all possible stereochemically isomeric forms.

Stereocentres are drawn in the usual style using dashed lines or wedge lines. E.g:

Boc-N-methylalanine (S)-(+)-2-hydroxy-2-phenylpropanoic acid

When the compound is a mixture of two non-mirromeric isomers / epi isomers, the configuration of the stereocenter is not particularly pointed out, but is represented by a straight line.

Unless otherwise mentioned or stated, when a compound of formula (I) contains one or more chiral centres, it may exist as two or more optical isomers. Citations include, unless otherwise specified in the specification, all of their optical isomer chemical formulas (ie, palm isomers, epimers, and non-image isomers) may also be individual optical isomers or A mixture (e.g., a racemic mixture) or two or more optical isomers.

Optical isomers can be characterized and identified by their optical activity (ie, + and-isomers, or d and l isomers), or they can be expressed in their absolute stereochemistry using the "R and S" nomenclature. The nomenclature was developed by Cahn, Ingold, and Prelog. See Advanced Organic Chemistry by Jerry March, 4th Edition, John Wiley & Sons, New York, 1992, pages 109-114 and Cahn, Ingold & Prelog, Angew. Chem. Int. Ed. Engl., 1966, 5, 385-415.

Optical isomers can be separated by several techniques, including chiral chromatography (column chromatography using palm supports), such techniques are known to those skilled in the art.

In addition to palm chromatography, optical isomers can also be formed by forming a non-diastereoisomer salt with palmitic acid, and then separating the non-image isomer salt by a preferred crystallization method, and then dissociating the salt. The individual mirror isomers of the free base are separated, in which palmitic acids are (+)-tartaric acid, (-)-pyroglutamic acid, (-)-di-toluene Formyl-L-tartaric acid ((-)-di-toluoyl-L-tartaric acid), (+)-mandelic acid, (-)-malic acid, and (-)- Camphorsulfonic.

In addition, it is also possible to separate non-mirror isomers by covalently bonding a pure palmitative adjuvant with a mirror image isomer and then using conventional methods such as chromatography to achieve mirror image isomer separation. . Then, the aforementioned covalent bond is cleaved to obtain a suitable pure mirror image isomer product.

When the compound of formula (I) exists as two or more optical isomers, one of a pair of mirror isomers can exhibit better characteristics than the other mirror isomer, such as biological activity. Therefore, in certain situations, one would like to use one of a pair of mirror isomers or one of a plurality of non-mirro isomers as a therapeutic agent. Accordingly, the present invention provides a composition comprising a compound of formula (I) having one or more chiral centers, wherein at least 55% (e.g., at least 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%) are single optical isomers (eg, mirror image isomers or non-image isomers). In a general embodiment, 99% or more (e.g., substantially all) of the total compounds of formula (I) may be a single optical isomer (e.g., a mirror image isomer or a non-image isomer物).

Compounds containing double bonds may have an E (anti-side) or Z (i-side) stereochemistry at the double bond. The substituent on the bivalent ring or (partially) saturated radical may be one of the cis or trans configuration. The terms cis and trans used herein are according to Chemical Abstracts nomenclature (J. Org. Chem. 1970, 35 (9), 2849-2867) and refer to the position of the substituent on the cyclic group.

Of particular interest are those whose compounds of formula (I) have pure stereochemical properties. For example, when a compound of formula (I) is designated by R, it means that the compound does not substantially include an S-type isomer. If a compound of formula (I) is designated by E, it means that the compound does not substantially include the Z-isomer. Ci, trans, R, S, E and Z are all words known in the art.

Isotope change

The present invention includes all pharmaceutically acceptable isotopically-labeled compounds of the present invention, that is, one or more atoms in the compound of formula (I) have the same atomic order but their atomic weight or mass number is generally the same as that found in nature. Number of different atoms.

Examples of isotopes suitable for the compounds of the present invention include: hydrogen isotopes, such as ² H (D) and ³ H (T); carbon isotopes, such as ¹¹ C, ¹³ C, and ¹⁴ C; isotopes of chlorine, such as ³⁶ Cl; fluorine Isotopes such as ¹⁸ F; isotopes of iodine such as ¹²³ I, ¹²⁵ I and ¹³¹ I; isotopes of nitrogen such as ¹³ N and ¹⁵ N; isotopes of oxygen such as ¹⁵ O, ¹⁷ O and ¹⁸ O; isotopes of phosphorus, Such as ³² P; and isotopes of sulfur, such as ³⁵ S.

The compounds of formula (I) calibrated by specific isotopes, such as those containing radioisotopes, can be applied to the study of the distribution of drugs and / or tissues. The compound of formula (I) can also have valuable diagnostic properties, which can be used to detect or determine the formation of a calibration compound and other molecules, peptides, proteins, enzymes or receptor complexes. Detection or calibration methods can use compounds calibrated with calibration reagents, where the calibration reagents can be radioisotopes, enzymes, fluorescent substances, chemiluminescent substances (e.g., luminol, luminescent amine derivatives, luciferin ), Aequorin, and luciferase). The radioisotope tritium (ie, ³ H (T)) and carbon-14 (ie, ¹⁴ C) are particularly suitable for this purpose because they are easy to use and common detection methods.

Substituents with heavier isotopes, such as deuterium (ie, ² H (D)) can also have specific therapeutic advantages, resulting in greater metabolic stability. For example, its half-life in vivo can increase or decrease the dose requirement, so it is applicable. In several situations.

Substituents with positive positron isotopes, such as ¹¹ C, ¹⁸ F, ¹⁵ O, and ¹³ N, can be used in positron emission tomography (PET) studies to detect target occupancy.

Isotopically-calibrated compounds of formula (I) can generally be prepared by conventional techniques known in the art, or by similar processes to the examples or preparations, but using appropriate isotopic calibration reagents instead of the non-calibration reagents used above.

ester

Compounds of formula (I), such as carboxylic acid esters or hydroxyl-containing esters, such as carboxylic acid esters, acyloxy esters and phosphate esters, are also included in chemical formula (I). Examples of esters include compounds containing a -C (= O) OR group, where R is an ester substituent, such as a C _1-7 alkyl group, a C _3-12 heterocyclic group, or a C _5-12 aryl group, preferably C _1-6 alkyl. Specific examples of ester groups include, but are not limited to, -C (= O) OCH ₃ , -C (= O) OCH ₂ CH ₃ , -C (= O) OC (CH ₃ ) ₃ and -C (= O) OPh. Examples of fluorenyl (reverse ester) groups are shown in -OC (= O) R, where R is a fluorenyl substituent, such as C _1-6 alkyl, C _3-12 heterocyclyl, or C _{5 -12} aryl, preferably C _1-6 alkyl. Specific examples of alkoxy include, but are not limited to, -OC (= O) CH ₃ (ethoxy), -OC (= O) CH ₂ CH ₃ , -OC (= O) C (CH ₃ ) ₃ , -OC (= O) Ph, and -OC (= O) CH ₂ Ph. Examples of phosphates are derived from phosphoric acid.

In a specific embodiment of the present invention, the chemical formula (I) includes the scope of the compound of the chemical formula (I) containing carboxylic acid group or hydroxyl ester. In another specific embodiment of the present invention, the formula (I) does not include the scope of the compound of the formula (I) containing carboxylic acid groups or hydroxyl-containing esters.

Hydrate and crystalline form

Chemical formula (I) also includes any polymorphic form of the compound, as well as solvates such as hydrates, alcoholates, and the like.

The compounds of the present invention may form solvates, for example, may form solvates with water (ie, hydrates) or common organic solvents. As used herein, the term "solvate" refers to the physical association of a compound of the invention with one or more solvent molecules. This physical association involves varying degrees of ionic and covalent bonding, including hydrogen bonding. Solvates can be separated under specific circumstances, for example, when one or more solvent molecules are incorporated in the crystal lattice of a crystalline solid. The term "solvate" is also meant to include both solution-phase and separable solvates. Non-limiting examples of suitable solvates include: the compounds of the present invention in combination with water, isopropanol, ethanol, methanol, DMSO, ethyl acetate, acetic acid or ethanolamine, and the like. The compounds of the present invention can exhibit their physiological effects despite being in a solvent.

Solvates are widely known in medicinal chemistry. Solvent compounds are important for the manufacturing process of a substance (e.g., related to its purification), storage of the substance (e.g., its stability), and ease of operation of the substance, and solvent compounds are often part of the isolation or purification stage of chemical synthesis. Those skilled in the art to which the present invention pertains may determine whether the separation conditions or purification conditions used have hydrates or other solvates formed according to standard and long-term technical means to prepare a predetermined compound. Such technologies include thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), X-ray crystal analysis (e.g., single crystal X-ray crystal analysis or X-ray powder diffraction), and solid state NMR (SS-NMR, also Called Magic Angle Spinning (NMR or MAS-NMR). Such techniques can also be part of standard analytical tools known to those skilled in the art, such as NMR, IR, HPLC, and MS.

On the other hand, those skilled in the art can use crystallization conditions to deliberately form a solvate, wherein the crystallization conditions include an amount of a solvent for forming a specific solvate. Thereafter, the standard methods described above can be used to determine whether a solvate is formed.

In a specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3 , 2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} piperazine -1-yl] ethan-1-one salt has <10% solvate present (for example, does not exceed the following amount: 9, 8, 7, 6, 5, 4, 3, 2, 1, 0.5, 0.1 (0.05, 0.01, or 0.01%), such as hydrates, alcoholates, isopropylacetate, methyl acetate, or alkanes, such as heptanes.

In a specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3 , 2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} piperazine The salt of 1-1-yl] ethyl-1-one is anhydrous. In yet another embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [ 3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} piperazine Anhydrous salt of azin-1-yl] ethyl-1-one contains not more than 5% by weight (for example, not more than any of the following amounts 4, 3, 2, 1, 0.5, 0.1, 0.05, or 0.01% ) Of water.

In a specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3 , 2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} piperazine The salt of -1-yl] ethyl-1-one contains a single bond crystalline form, and other crystalline forms do not exceed 5% by weight (for example, not more than 4, 3, 2, 1, 0.5, 0.1, 0.05, or 0.01% Either.

In a specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3 , 2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} piperazine The salt of 1-1-yl] ethyl-1-one is crystalline.

In a specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3 , 2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} piperazine The salt of 1-1-yl] ethyl-1-one is amorphous.

Furthermore, the compounds of the present invention may have one or more polymorph or amorphous crystalline forms, and they are included in the scope of the invention.

As used herein, "polymorph" means that the compound of formula (I) has more than one crystalline structure. The ability of a chemical compound to crystallize into more than one crystal form can have an effect on the properties of the compound, such as physical and chemical properties, shelf life, solubility, formulation properties, toxicity, bioavailability, water absorption, and processability . In addition, the therapeutic effects of pharmaceutical compounds can be affected by the polymorphism of drug molecules.

In a specific embodiment, the compound of formula (I) includes 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl ] Methyl} piperazin-1-yl] polymorphs of ethyl-1-one or a salt thereof.

In another specific embodiment, the compound of formula (I) includes 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H , 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholine-4- Polymorphs of the salt of [methyl] methyl} piperazin-1-yl] ethan-1-one.

In another specific embodiment, the compound of formula (I) includes 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H , 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholine-4- Form A polymorph of methyl] methyl} piperazin-1-yl] ethyl-1-one L-(+)-lactate. This compound can be prepared as described in Example 39 herein.

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} Piperazin-1-yl] ethyl-1-one L-(+)-lactate (1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H , 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[(3R) -3-methylmorpholin-4-yl] methyl} piperazin The ¹ H NMR spectral characteristics of -1-yl] ethan-1-one L-(+)-lactate) are shown in FIG. 1.

The X-ray powder diffraction pattern of the compound is characterized by the diffraction angle (2θ) and the interplanar spacing (d) of the X-ray diffraction spectrum. These are related to the following Bragg's equation: nλ = 2d Sin θ, (here, n = 1; λ = wavelength of the cathode used; d = plane spacing; and θ = diffraction angle). In this paper, the crystal plane spacing, diffraction angle, and overall pattern are important for crystal identification in X-ray powder diffraction, which is due to the characteristics of the data. Because the relative strength varies with the crystal growth direction, particle size, and measurement conditions, it should not be interpreted strictly. In addition, the diffraction angle generally refers to an angle that conforms to the angle 2θ ± 0.2 °. The crest refers to the main crest and includes crests whose diffraction angles other than the above are not greater than the median.

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} Piperazin-1-yl] ethyl-1-one L-(+)-lactate is a type A polymorph with XRPD pattern peak positions as follows: 6.5 ± 0.5 °, 7.1 ± 0.5 °, 7.9 ± 0.5 °, 9.3 ± 0.5 °, 10.2 ± 0.5 °, 11.0 ± 0.5 °, 11.6 ± 0.5 °, 13.3 ± 0.5 °, 14.4 ± 0.5 °, 15.0 ± 0.5 °, 16.7 ± 0.5 °, 18.0 ± 0.5 °, 18.4 ± 0.5 °, 20.0 ± 0.5 °, 21.0 ± 0.5 °, 23.4 ± 0.5 °, 25.2 ± 0.5 ° and 26.1 ± 0.5 ° (2θ, 1d.p).

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} Piperazin-1-yl] ethyl-1-one L-(+)-lactate type A polymorph is characterized by having XRPD pattern peak positions as follows: 6.5 ± 0.2 °, 7.1 ± 0.2 °, 7.9 ± 0.2 °, 9.3 ± 0.2 °, 10.2 ± 0.2 °, 11.0 ± 0.2 °, 11.6 ± 0.2 °, 13.3 ± 0.2 °, 14.4 ± 0.2 °, 15.0 ± 0.2 °, 16.7 ± 0.2 °, 18.0 ± 0.2 °, 18.4 ± 0.2 °, 20.0 ± 0.2 °, 21.0 ± 0.2 °, 23.4 ± 0.2 °, 25.2 ± 0.2 ° and 26.1 ± 0.2 ° (2θ, 1d.p).

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} Piperazin-1-yl] ethyl-1-one L-(+)-lactate type A polymorph is characterized by having XRPD pattern peak positions as follows: 6.5 ± 0.1 °, 7.1 ± 0.1 °, 7.9 ± 0.1 °, 9.3 ± 0.1 °, 10.2 ± 0.1 °, 11.0 ± 0.1 °, 11.6 ± 0.1 °, 13.3 ± 0.1 °, 14.4 ± 0.1 °, 15.0 ± 0.1 °, 16.7 ± 0.1 °, 18.0 ± 0.1 °, 18.4 ± 0.1 °, 20.0 ± 0.1 °, 21.0 ± 0.1 °, 23.4 ± 0.1 °, 25.2 ± 0.1 °, and 26.1 ± 0.1 ° (2θ, 1d.p).

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} Piperazin-1-yl] ethan-1-one L-(+)-lactate type A polymorph is characterized by having XRPD pattern peak positions as follows: 6.5 °, 7.1 °, 7.9 °, 9.3 °, 10.2 °, 11.0 °, 11.6 °, 13.3 °, 14.4 °, 15.0 °, 16.7 °, 18.0 °, 18.4 °, 20.0 °, 21.0 °, 23.4 °, 25.2 ° and 26.1 ° (2θ, 1d.p).

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} The XRPD pattern characteristics of the piperazin-1-yl] ethyl-1-one L-(+)-lactate type A polymorph are substantially shown in FIG. 2.

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} The piperazin-1-yl] ethyl-1-one L-(+)-lactate type A polymorph is characterized by a peak having the same diffraction angle (2θ) as the XRPD pattern in FIG. 2 and is selective, These peaks have the same relative intensity as the peaks in FIG. 2.

Those skilled in the art should know that the "intensity" of the XRPD peak referred to herein refers to the relative intensity, which has taken into account the background noise and the normalisation of these other parameters.

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} The piperazin-1-yl] ethyl-1-one L-(+)-lactate type A polymorph is characterized by having a diffraction angle (2θ) and intensity of the main peak shown in the XRPD pattern in FIG. 2.

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} Piperazin-1-yl] ethyl-1-one L-(+)-lactate type A polymorph is characterized by its interplanar spacing (d) values: 13.59 ± 0.5Å, 12.44 ± 0.5Å, 11.19 ± 0.5Å, 9.50 ± 0.5Å, 8.67 ± 0.5Å, 8.04 ± 0.5Å, 7.62 ± 0.5Å, 6.65 ± 0.5Å, 6.15 ± 0.5Å, 5.90 ± 0.5Å, 5.31 ± 0.5Å, 4.93 ± 0.5Å, 4.82 ± 0.5Å, 4.44 ± 0.5Å, 4.23 ± 0.5Å, 3.80 ± 0.5Å, 3.53 ± 0.5Å, and 3.41 ± 0.5Å (d, 2d.p.).

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} Piperazin-1-yl] ethyl-1-one L-(+)-lactate type A polymorph is characterized by its interplanar spacing (d) values: 13.59 ± 0.2Å, 12.44 ± 0.2Å, 11.19 ± 0.2Å, 9.50 ± 0.2Å, 8.67 ± 0.2Å, 8.04 ± 0.2Å, 7.62 ± 0.2Å, 6.65 ± 0.2Å, 6.15 ± 0.2Å, 5.90 ± 0.2Å, 5.31 ± 0.2Å, 4.93 ± 0.2Å, 4.82 ± 0.2Å, 4.44 ± 0.2Å, 4.23 ± 0.2Å, 3.80 ± 0.2Å, 3.53 ± 0.2Å, and 3.41 ± 0.2Å (d, 2d.p.).

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} Piperazin-1-yl] ethyl-1-one L-(+)-lactate type A polymorph is characterized by its interplanar spacing (d) values: 13.59 ± 0.1Å, 12.44 ± 0.1Å, 11.19 ± 0.1Å, 9.50 ± 0.1Å, 8.67 ± 0.1Å, 8.04 ± 0.1Å, 7.62 ± 0.1Å, 6.65 ± 0.1Å, 6.15 ± 0.1Å, 5.90 ± 0.1Å, 5.31 ± 0.1Å, 4.93 ± 0.1Å, 4.82 ± 0.1Å, 4.44 ± 0.1Å, 4.23 ± 0.1Å, 3.80 ± 0.1Å, 3.53 ± 0.1Å and 3.41 ± 0.1Å (d, 2d.p.).

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} Piperazin-1-yl] ethyl-1-one L-(+)-lactate type A polymorph is characterized by its interplanar spacing (d) values: 13.59Å, 12.44Å, 11.19Å, 9.50Å, 8.67Å, 8.04Å, 7.62Å, 6.65Å, 6.15Å, 5.90Å, 5.31Å, 4.93Å, 4.82Å, 4.44Å, 4.23Å, 3.80Å, 3.53Å and 3.41Å (d, 2d.p.).

In yet another embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [ 3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} piperazine Azine-1-yl] ethyl-1-one L-(+)-lactate type A polymorphs are characterized by a DSC peak temperature of 78.69ºC ± 0.5ºC and / or 113.91ºC ± 0.5ºC (eg 78.69ºC ± 0.2ºC and / or 113.91ºC ± 0.2ºC, especially 78.69ºC ± 0.1ºC and / or 113.91ºC ± 0.1ºC, and more specifically 78.69ºC and / or 113.91ºC).

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} Piperazin-1-yl] ethyl-1-one L-(+)-lactate type A polymorphs are characterized by a DSC onset temperature of 72.3ºC ± 0.5ºC (endothermic, broad wave) and / or 102ºC ± 0.5 ºC (endothermic, wide wave) (eg 72.3ºC ± 0.2ºC and / or 102ºC ± 0.2ºC, especially 72.3ºC ± 0.1ºC and / or 102ºC ± 0.1ºC, and more particularly 72.3ºC and / or 102ºC).

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} The piperazin-1-yl] ethyl-1-one L-(+)-lactate type A polymorph is characterized by a DSC thermogram as shown in FIG. 3.

In another specific embodiment, the compound of formula (I) includes 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H , 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholine-4- [Formula] methyl] piperazin-1-yl] ethan-1-one L-(+)-lactate B-type polymorph. This compound can be prepared as described in Example 40 herein.

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} The ¹ H NMR spectral characteristics of piperazin-1-yl] ethyl-1-one L-(+)-lactate are shown in FIG. 4.

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} Piperazin-1-yl] ethyl-1-one L-(+)-lactate type B polymorph is characterized by having XRPD pattern peak positions as follows: 6.6 ± 0.5 °, 9.4 ± 0.5 °, 11.0 ± 0.5 °, 13.2 ± 0.5 °, 14.3 ± 0.5 °, 15.8 ± 0.5 °, 17.4 ± 0.5 °, 18.4 ± 0.5 °, 19.1 ± 0.5 °, 20.9 ± 0.5 °, 21.8 ± 0.5 °, 23.1 ± 0.5 °, 24.9 ± 0.5 °, 26.7 ± 0.5 ° and 27.8 ± 0.5 ° (2θ, 1d.p).

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} Piperazin-1-yl] ethyl-1-one L-(+)-lactate type B polymorphs are characterized by having XRPD pattern peak positions as follows: 6.6 ± 0.2 °, 9.4 ± 0.2 °, 11.0 ± 0.2 °, 13.2 ± 0.2 °, 14.3 ± 0.2 °, 15.8 ± 0.2 °, 17.4 ± 0.2 °, 18.4 ± 0.2 °, 19.1 ± 0.2 °, 20.9 ± 0.2 °, 21.8 ± 0.2 °, 23.1 ± 0.2 °, 24.9 ± 0.2 °, 26.7 ± 0.2 ° and 27.8 ± 0.2 ° (2θ, 1d.p).

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} Piperazin-1-yl] ethyl-1-one L-(+)-lactate type B polymorphs are characterized by having XRPD pattern peak positions as follows: 6.6 ± 0.1 °, 9.4 ± 0.1 °, 11.0 ± 0.1 °, 13.2 ± 0.1 °, 14.3 ± 0.1 °, 15.8 ± 0.1 °, 17.4 ± 0.1 °, 18.4 ± 0.1 °, 19.1 ± 0.1 °, 20.9 ± 0.1 °, 21.8 ± 0.1 °, 23.1 ± 0.1 °, 24.9 ± 0.1 °, 26.7 ± 0.1 ° and 27.8 ± 0.1 ° (2θ, 1d.p).

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} Piperazin-1-yl] ethyl-1-one L-(+)-lactate type B polymorphs are characterized by having XRPD pattern peak positions as follows: 6.6 °, 9.4 °, 11.0 °, 13.2 °, 14.3 °, 15.8 °, 17.4 °, 18.4 °, 19.1 °, 20.9 °, 21.8 °, 23.1 °, 24.9 °, 26.7 ° and 27.8 ° (2θ, 1d.p).

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} The XRPD pattern characteristics of the piperazin-1-yl] ethyl-1-one L-(+)-lactate B-type polymorph are substantially as shown in FIG. 5.

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} Piperazin-1-yl] ethyl-1-one L-(+)-lactate type B polymorph is characterized by a peak having the same diffraction angle (2θ) as the XRPD pattern in FIG. 5, and selective, These peaks have the same relative intensity as the peaks in FIG. 5.

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} The B-type polymorph of piperazin-1-yl] ethyl-1-one L-(+)-lactate is characterized by the diffraction angle (2θ) and intensity of the main peak shown in the XRPD pattern in FIG. 5.

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} Piperazin-1-yl] ethyl-1-one L-(+)-lactate type B polymorph is characterized by its interplanar spacing (d) values: 13.39 ± 0.5Å, 9.40 ± 0.5Å, 8.04 ± 0.5Å, 6.70 ± 0.5Å, 6.19 ± 0.5Å, 5.61 ± 0.5Å, 5.09 ± 0.5Å, 4.82 ± 0.5Å, 4.64 ± 0.5Å, 4.25 ± 0.5Å, 4.07 ± 0.5Å, 3.85 ± 0.5Å, 3.57 ± 0.5Å, 3.34 ± 0.5Å and 3.21 ± 0.5Å (d, 2d.p.).

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} Piperazin-1-yl] ethyl-1-one L-(+)-lactate type B polymorph is characterized by its interplanar spacing (d) values: 13.39 ± 0.2Å, 9.40 ± 0.2Å, 8.04 ± 0.2Å, 6.70 ± 0.2Å, 6.19 ± 0.2Å, 5.61 ± 0.2Å, 5.09 ± 0.2Å, 4.82 ± 0.2Å, 4.64 ± 0.2Å, 4.25 ± 0.2Å, 4.07 ± 0.2Å, 3.85 ± 0.2Å, 3.57 ± 0.2Å, 3.34 ± 0.2Å and 3.21 ± 0.2Å (d, 2d.p.).

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} Piperazin-1-yl] ethyl-1-one L-(+)-lactate type B polymorph is characterized by its interplanar spacing (d) values: 13.39 ± 0.1Å, 9.40 ± 0.1Å, 8.04 ± 0.1Å, 6.70 ± 0.1Å, 6.19 ± 0.1Å, 5.61 ± 0.1Å, 5.09 ± 0.1Å, 4.82 ± 0.1Å, 4.64 ± 0.1Å, 4.25 ± 0.1Å, 4.07 ± 0.1Å, 3.85 ± 0.1Å, 3.57 ± 0.1Å, 3.34 ± 0.1Å and 3.21 ± 0.1Å (d, 2d.p.).

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} Piperazin-1-yl] ethyl-1-one L-(+)-lactate type B polymorph is characterized by its interplanar spacing (d) values: 13.39Å, 9.40Å, 8.04Å, 6.70Å, 6.19Å, 5.61Å, 5.09Å, 4.82Å, 4.64Å, 4.25Å, 4.07Å, 3.85Å, 3.57Å, 3.34Å and 3.21Å (d, 2d.p.).

In yet another embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [ 3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} piperazine Azine-1-yl] ethyl-1-one L-(+)-lactate type B polymorph is characterized by a DSC peak temperature of 85.25ºC ± 0.5ºC and / or 106.72ºC ± 0.5ºC (such as 85.25ºC ± 0.2 ºC and / or 106.72ºC ± 0.2ºC, especially 85.25ºC ± 0.1ºC and / or 106.72ºC ± 0.1ºC, and more specifically 85.25ºC and / or 106.72ºC).

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} Piperazin-1-yl] ethyl-1-one L-(+)-lactate type B polymorphs are characterized by a DSC onset temperature of 68ºC ± 0.5ºC (large endothermic, broad wave) and / or 102ºC ± 0.5 ºC (small endothermic, wide wave) (eg 68ºC ± 0.2ºC and / or 102ºC ± 0.2ºC, especially 68ºC ± 0.1ºC and / or 102ºC ± 0.1ºC, more specifically 68ºC and / or 102ºC).

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} The DSC temperature diagram of piperazin-1-yl] ethyl-1-one L-(+)-lactate B-type polymorph is shown in FIG. 6.

In another specific embodiment, the compound of formula (I) includes 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H , 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholine-4- [Formula] methyl] piperazin-1-yl] ethan-1-one sulfate F polymorph. This compound can be prepared as described in Example 41 herein.

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} The ¹ H NMR spectrum characteristics of piperazin-1-yl] ethyl-1-one sulfate are shown in FIG. 7.

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} Piperazin-1-yl] ethyl-1-one sulfate F-type polymorphs are characterized by having XRPD pattern peak positions as follows: 8.5 ± 0.5 °, 13.5 ± 0.5 °, 13.9 ± 0.5 °, 14.3 ± 0.5 °, 16.2 ± 0.5 °, 17.3 ± 0.5 °, 20.1 ± 0.5 °, 21.3 ± 0.5 °, 23.3 ± 0.5 °, 24.4 ± 0.5 °, and 27.9 ± 0.5 ° (2θ, 1d.p).

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} Piperazin-1-yl] ethyl-1-one sulfate F-type polymorphs are characterized by having XRPD pattern peak positions as follows: 8.5 ± 0.2 °, 13.5 ± 0.2 °, 13.9 ± 0.2 °, 14.3 ± 0.2 °, 16.2 ± 0.2 °, 17.3 ± 0.2 °, 20.1 ± 0.2 °, 21.3 ± 0.2 °, 23.3 ± 0.2 °, 24.4 ± 0.2 °, and 27.9 ± 0.2 ° (2θ, 1d.p).

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} Piperazin-1-yl] ethyl-1-one sulfate F-type polymorphs are characterized by having XRPD pattern peak positions as follows: 8.5 ± 0.1 °, 13.5 ± 0.1 °, 13.9 ± 0.1 °, 14.3 ± 0.1 °, 16.2 ± 0.1 °, 17.3 ± 0.1 °, 20.1 ± 0.1 °, 21.3 ± 0.1 °, 23.3 ± 0.1 °, 24.4 ± 0.1 ° and 27.9 ± 0.1 ° (2θ, 1d.p).

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} Piperazin-1-yl] ethyl-1-one sulfate F-type polymorphs are characterized by having XRPD pattern peak positions as follows: 8.5 °, 13.5 °, 13.9 °, 14.3 °, 16.2 °, 17.3 °, 20.1 °, 21.3 °, 23.3 °, 24.4 ° and 27.9 ° (2θ, 1d.p).

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} The XRPD pattern characteristics of the F-type polymorph of piperazin-1-yl] ethyl-1-one sulfate are shown in FIG. 8.

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} Piperazin-1-yl] ethyl-1-one sulfate F-type polymorphs are characterized by peaks having the same diffraction angle (2θ) as the XRPD pattern in FIG. 8 and are selective, in which these peaks have The peaks in Figure 8 have the same relative intensity.

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} The F-type polymorph of piperazin-1-yl] ethan-1-one sulfate is characterized by having a diffraction angle (2θ) and an intensity of the main peak shown in the XRPD pattern in FIG. 8.

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} The F-type polymorph of piperazin-1-yl] ethyl-1-one sulfate is characterized by its interplanar spacing (d) values: 10.40 ± 0.5Å, 6.56 ± 0.5Å, 6.37 ± 0.5Å, 6.19 ± 0.5 Å, 5.47 ± 0.5Å, 5.12 ± 0.5Å, 4.42 ± 0.5Å, 4.17 ± 0.5Å, 3.82 ± 0.5Å, 3.65 ± 0.5Å and 3.20 ± 0.5Å (d, 2d.p.).

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} Piperazin-1-yl] ethyl-1-one sulfate F-type polymorphs are characterized by their interplanar spacing (d) values: 10.40 ± 0.2Å, 6.56 ± 0.2Å, 6.37 ± 0.2Å, 6.19 ± 0.2 Å, 5.47 ± 0.2Å, 5.12 ± 0.2Å, 4.42 ± 0.2Å, 4.17 ± 0.2Å, 3.82 ± 0.2Å, 3.65 ± 0.2Å and 3.20 ± 0.2Å (d, 2d.p.).

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} The F-type polymorph of piperazin-1-yl] ethyl-1-one sulfate is characterized by its interplanar spacing (d) values: 10.40 ± 0.1Å, 6.56 ± 0.1Å, 6.37 ± 0.1Å, 6.19 ± 0.1 Å, 5.47 ± 0.1Å, 5.12 ± 0.1Å, 4.42 ± 0.1Å, 4.17 ± 0.1Å, 3.82 ± 0.1Å, 3.65 ± 0.1Å and 3.20 ± 0.1Å (d, 2d.p.).

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} Piperazin-1-yl] ethyl-1-one sulfate F-type polymorphs are characterized by interplanar spacing (d) values: 10.40Å, 6.56Å, 6.37Å, 6.19Å, 5.47Å, 5.12Å, 4.42Å, 4.17Å, 3.82Å, 3.65Å and 3.20Å (d, 2d.p.).

In yet another embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [ 3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} piperazine The F-type polymorph of azin-1-yl] ethyl-1-one sulfate is characterized by a DSC peak temperature of 80.31ºC ± 0.5ºC and / or 149.07ºC ± 0.5ºC (eg, 80.31ºC ± 0.2ºC and / or 149.07ºC ± 0.2ºC, especially 80.31ºC ± 0.1ºC and / or 149.07ºC ± 0.1ºC, more specifically 80.31ºC and / or 149.07ºC).

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} Piperazin-1-yl] ethyl-1-one sulfate F-type polymorphs are characterized by a DSC onset temperature of 51.2ºC ± 0.5ºC (endothermic, broad peak) and / or 136ºC ± 0.5ºC (endothermic, broad peak ) (Eg 51.2ºC ± 0.2ºC and / or 136ºC ± 0.2ºC, especially 51.2ºC ± 0.1ºC and / or 136ºC ± 0.1ºC, more specifically 51.2ºC and / or 136ºC).

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} The characteristics of the DSC temperature chart of the F-type polymorph of piperazin-1-yl] ethyl-1-one sulfate are shown in FIG. 9.

In another specific embodiment, the compound of formula (I) includes 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H , 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholine-4- [Formula] methyl] piperazin-1-yl] ethyl-1-ketomethanesulfonate polymorph. This compound can be prepared as described in Example 42 herein.

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} The ¹ H NMR spectrum characteristic of piperazin-1-yl] ethyl-1-one mesylate is shown in FIG. 10.

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} Piperazin-1-yl] ethyl-1-one mesylate type B polymorph is characterized by having XRPD pattern peak positions as follows: 6.6 ± 0.5 °, 8.0 ± 0.5 °, 11.8 ± 0.5 °, 13.2 ± 0.5 ° , 14.3 ± 0.5 °, 15.0 ± 0.5 °, 15.6 ± 0.5 °, 17.1 ± 0.5 °, 17.4 ± 0.5 °, 17.7 ± 0.5 °, 19.2 ± 0.5 °, 20.3 ± 0.5 °, 21.2 ± 0.5 °, 22.3 ± 0.5 ° , 23.0 ± 0.5 °, 24.0 ± 0.5 °, 25.8 ± 0.5 °, 26.8 ± 0.5 °, and 28.9 ± 0.5 ° (2θ, 1d.p).

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} Piperazin-1-yl] ethyl-1-one mesylate type B polymorphs are characterized by having XRPD pattern peak positions as follows: 6.6 ± 0.2 °, 8.0 ± 0.2 °, 11.8 ± 0.2 °, 13.2 ± 0.2 ° , 14.3 ± 0.2 °, 15.0 ± 0.2 °, 15.6 ± 0.2 °, 17.1 ± 0.2 °, 17.4 ± 0.2 °, 17.7 ± 0.2 °, 19.2 ± 0.2 °, 20.3 ± 0.2 °, 21.2 ± 0.2 °, 22.3 ± 0.2 ° , 23.0 ± 0.2 °, 24.0 ± 0.2 °, 25.8 ± 0.2 °, 26.8 ± 0.2 ° and 28.9 ± 0.2 ° (2θ, 1d.p).

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} Piperazin-1-yl] ethyl-1-one mesylate type B polymorphs are characterized by having XRPD pattern peak positions as follows: 6.6 ± 0.1 °, 8.0 ± 0.1 °, 11.8 ± 0.1 °, 13.2 ± 0.1 ° , 14.3 ± 0.1 °, 15.0 ± 0.1 °, 15.6 ± 0.1 °, 17.1 ± 0.1 °, 17.4 ± 0.1 °, 17.7 ± 0.1 °, 19.2 ± 0.1 °, 20.3 ± 0.1 °, 21.2 ± 0.1 °, 22.3 ± 0.1 ° , 23.0 ± 0.1 °, 24.0 ± 0.1 °, 25.8 ± 0.1 °, 26.8 ± 0.1 °, and 28.9 ± 0.1 ° (2θ, 1d.p).

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} Piperazin-1-yl] ethyl-1-one mesylate type B polymorphs are characterized by having XRPD pattern peak positions as follows: 6.6 °, 8.0 °, 11.8 °, 13.2 °, 14.3 °, 15.0 °, 15.6 °, 17.1 °, 17.4 °, 17.7 °, 19.2 °, 20.3 °, 21.2 °, 22.3 °, 23.0 °, 24.0 °, 25.8 °, 26.8 °, and 28.9 ° (2θ, 1d.p).

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} The XRPD pattern characteristics of the B-type polymorph of piperazin-1-yl] ethan-1-one mesylate are shown in FIG. 11.

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} Piperazin-1-yl] ethan-1-one mesylate type B polymorph is characterized by a peak having the same diffraction angle (2θ) as the XRPD pattern in FIG. 11 and is selective, among which these peaks It has the same relative intensity as the peak in FIG. 11.

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} The B-type polymorph of piperazin-1-yl] ethan-1-one mesylate is characterized by the diffraction angle (2θ) and the intensity of the main peak shown in the XRPD pattern in FIG. 11.

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} Piperazin-1-yl] ethyl-1-one mesylate type B polymorph is characterized by its interplanar spacing (d) values: 13.39 ± 0.5Å, 11.05 ± 0.5Å, 7.50 ± 0.5Å, 6.70 ± 0.5Å, 6.19 ± 0.5Å, 5.90 ± 0.5Å, 5.68 ± 0.5Å, 5.18 ± 0.5Å, 5.09 ± 0.5Å, 5.01 ± 0.5Å, 4.62 ± 0.5Å, 4.37 ± 0.5Å, 4.19 ± 0.5Å, 3.98 ± 0.5Å, 3.86 ± 0.5Å, 3.71 ± 0.5Å, 3.45 ± 0.5Å, 3.32 ± 0.5Å and 3.09 ± 0.5Å (d, 2d.p.).

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} Piperazin-1-yl] ethyl-1-one mesylate type B polymorph is characterized by its interplanar spacing (d) values: 13.39 ± 0.2Å, 11.05 ± 0.2Å, 7.50 ± 0.2Å, 6.70 ± 0.2Å, 6.19 ± 0.2Å, 5.90 ± 0.2Å, 5.68 ± 0.2Å, 5.18 ± 0.2Å, 5.09 ± 0.2Å, 5.01 ± 0.2Å, 4.62 ± 0.2Å, 4.37 ± 0.2Å, 4.19 ± 0.2Å, 3.98 ± 0.2Å, 3.86 ± 0.2Å, 3.71 ± 0.2Å, 3.45 ± 0.2Å, 3.32 ± 0.2Å and 3.09 ± 0.2Å (d, 2d.p.).

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} The piperazin-1-yl] ethyl-1-one mesylate type B polymorph is characterized by its interplanar spacing (d) values: 13.39 ± 0.1Å, 11.05 ± 0.1Å, 7.50 ± 0.1Å, 6.70 ± 0.1Å, 6.19 ± 0.1Å, 5.90 ± 0.1Å, 5.68 ± 0.1Å, 5.18 ± 0.1Å, 5.09 ± 0.1Å, 5.01 ± 0.1Å, 4.62 ± 0.1Å, 4.37 ± 0.1Å, 4.19 ± 0.1Å, 3.98 ± 0.1Å, 3.86 ± 0.1Å, 3.71 ± 0.1Å, 3.45 ± 0.1Å, 3.32 ± 0.1Å, and 3.09 ± 0.1Å (d, 2d.p.).

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} The piperazine-1-yl] ethyl-1-one mesylate type B polymorph is characterized by its interplanar spacing (d) values: 13.39Å, 11.05Å, 7.50Å, 6.70Å, 6.19Å, 5.90 Å, 5.68Å, 5.18Å, 5.09Å, 5.01Å, 4.62Å, 4.37Å, 4.19Å, 3.98Å, 3.86Å, 3.71Å, 3.45Å, 3.32Å and 3.09Å (d, 2d.p.).

In yet another embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [ 3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} piperazine The B-type polymorph of azin-1-yl] ethyl-1-one mesylate is characterized by having a DSC peak temperature of 98.63ºC ± 0.5ºC and / or 177.11ºC ± 0.5ºC (for example, 98.63ºC ± 0.2ºC and / Or 177.11ºC ± 0.2ºC, especially 98.63ºC ± 0.1ºC and / or 177.11ºC ± 0.1ºC, more specifically 98.63ºC and / or 177.11ºC).

In yet another embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [ 3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} piperazine Azine-1-yl] ethyl-1-one mesylate type B polymorphs are characterized by having a DSC onset temperature of 73.3ºC ± 0.5ºC (endothermic, broad wave) and / or 160.8ºC ± 0.5ºC (endothermic , Wide wave peak) (eg 73.3ºC ± 0.2ºC and / or 160.8ºC ± 0.2ºC, especially 73.3ºC ± 0.1ºC and / or 160.8ºC ± 0.1ºC, and more particularly 73.3ºC and / or 160.8ºC).

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} The characteristics of the DSC temperature chart of the B-type polymorph of piperazin-1-yl] ethan-1-one mesylate are shown in FIG. 12.

In another specific embodiment, the compound of formula (I) includes 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H , 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholine-4- C] polymorph of methyl] methyl} piperazin-1-yl] ethyl-1-one L-(+)-lactate. This compound can be prepared as described in Example 43 herein.

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} The ¹ H NMR spectrum characteristic of piperazin-1-yl] ethyl-1-one L-(+)-lactate is shown in FIG. 15.

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} Piperazin-1-yl] ethyl-1-one L-(+)-lactate C-type polymorphs are characterized by having XRPD pattern peak positions as follows: 7.4 ± 0.5 °, 7.9 ± 0.5 °, 8.3 ± 0.5 °, 8.7 ± 0.5 °, 9.0 ± 0.5 °, 10.4 ± 0.5 °, 11.2 ± 0.5 °, 11.6 ± 0.5 °, 12.3 ± 0.5 °, 13.1 ± 0.5 °, 13.9 ± 0.5 °, 14.7 ± 0.5 °, 15.8 ± 0.5 °, 16.5 ± 0.5 °, 17.1 ± 0.5 °, 17.9 ± 0.5 °, 18.4 ± 0.5 °, 18.9 ± 0.5 °, 19.6 ± 0.5 °, 20.4 ± 0.5 °, 21.0 ± 0.5 °, 21.8 ± 0.5 °, 22.9 ± 0.5 °, 23.3 ± 0.5 °, 23.6 ± 0.5 °, 24.0 ± 0.5 °, 24.9 ± 0.5 °, and 26.4 ± 0.5 ° (2θ, 1d.p).

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} Piperazin-1-yl] ethyl-1-one L-(+)-lactate of the C-type polymorph is characterized by having XRPD pattern peak positions as follows: 7.4 ± 0.2 °, 7.9 ± 0.2 °, 8.3 ± 0.2 °, 8.7 ± 0.2 °, 9.0 ± 0.2 °, 10.4 ± 0.2 °, 11.2 ± 0.2 °, 11.6 ± 0.2 °, 12.3 ± 0.2 °, 13.1 ± 0.2 °, 13.9 ± 0.2 °, 14.7 ± 0.2 °, 15.8 ± 0.2 °, 16.5 ± 0.2 °, 17.1 ± 0.2 °, 17.9 ± 0.2 °, 18.4 ± 0.2 °, 18.9 ± 0.2 °, 19.6 ± 0.2 °, 20.4 ± 0.2 °, 21.0 ± 0.2 °, 21.8 ± 0.2 °, 22.9 ± 0.2 °, 23.3 ± 0.2 °, 23.6 ± 0.2 °, 24.0 ± 0.2 °, 24.9 ± 0.2 ° and 26.4 ± 0.2 ° (2θ, 1d.p).

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} Piperazin-1-yl] ethyl-1-one L-(+)-lactate of the C-type polymorph is characterized by having XRPD pattern peak positions as follows: 7.4 ± 0.1 °, 7.9 ± 0.1 °, 8.3 ± 0.1 °, 8.7 ± 0.1 °, 9.0 ± 0.1 °, 10.4 ± 0.1 °, 11.2 ± 0.1 °, 11.6 ± 0.1 °, 12.3 ± 0.1 °, 13.1 ± 0.1 °, 13.9 ± 0.1 °, 14.7 ± 0.1 °, 15.8 ± 0.1 °, 16.5 ± 0.1 °, 17.1 ± 0.1 °, 17.9 ± 0.1 °, 18.4 ± 0.1 °, 18.9 ± 0.1 °, 19.6 ± 0.1 °, 20.4 ± 0.1 °, 21.0 ± 0.1 °, 21.8 ± 0.1 °, 22.9 ± 0.1 °, 23.3 ± 0.1 °, 23.6 ± 0.1 °, 24.0 ± 0.1 °, 24.9 ± 0.1 ° and 26.4 ± 0.1 ° (2θ, 1d.p).

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} Piperazin-1-yl] ethan-1-one L-(+)-lactate C-type polymorphs are characterized by having XRPD pattern peak positions as follows: 7.4 °, 7.9 °, 8.3 °, 8.7 °, 9.0 °, 10.4 °, 11.2 °, 11.6 °, 12.3 °, 13.1 °, 13.9 °, 14.7 °, 15.8 °, 16.5 °, 17.1 °, 17.9 °, 18.4 °, 18.9 °, 19.6 °, 20.4 °, 21.0 °, 21.8 ° , 22.9 °, 23.3 °, 23.6 °, 24.0 °, 24.9 ° and 26.4 ° (2θ, 1d.p).

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} The XRPD pattern characteristics of piperazin-1-yl] ethyl-1-one L-(+)-lactate in the C-type polymorph are shown in FIG.

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} Piperazin-1-yl] ethyl-1-one L-(+)-lactate type C polymorph is characterized by a peak having the same diffraction angle (2θ) as the XRPD pattern labeled 1 in FIG. 16 and the selection In nature, these peaks have the same relative intensity as the peak labeled 1 in FIG. 16.

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} Piperazin-1-yl] ethyl-1-one L-(+)-lactate C-type polymorphs are characterized by having a diffraction angle (2θ) of the main peak as shown in the XRPD pattern labeled 1 in FIG. 16 And intensity.

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} Piperazin-1-yl] ethyl-1-one L-(+)-lactate type C polymorph is characterized by the main peaks measured by XRPD: 8.7 ± 0.5º, 17.1 ± 0.5 °, 17.9 ± 0.5 ° and 18.9 ± 0.5 ° (2θ, 1d.p).

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} Piperazin-1-yl] ethyl-1-one L-(+)-lactate type C polymorph is characterized by the main peaks measured by XRPD: 8.7 ± 0.2º, 17.1 ± 0.2 °, 17.9 ± 0.2 ° and 18.9 ± 0.2 ° (2θ, 1d.p).

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} Piperazin-1-yl] ethyl-1-one L-(+)-lactate type C polymorph is characterized by the main peaks measured by XRPD: 8.7 ± 0.1º, 17.1 ± 0.1 °, 17.9 ± 0.1 ° and 18.9 ± 0.1 ° (2θ, 1d.p).

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} Piperazin-1-yl] ethyl-1-one L-(+)-lactate C-type polymorphs are characterized by having the main peaks measured by XRPD: 8.7 °, 17.1 °, 17.9 ° and 18.9 ° ( 2θ, 1d.p).

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} The piperazine-1-yl] ethyl-1-one L-(+)-lactate C-type polymorph is characterized by its interplanar spacing (d) values: 11.94 ± 0.5Å, 11.19 ± 0.5Å, 10.65 ± 0.5Å, 10.16 ± 0.5Å, 9.82 ± 0.5Å, 8.50 ± 0.5Å, 7.90 ± 0.5Å, 7.62 ± 0.5Å, 7.19 ± 0.5Å, 6.75 ± 0.5Å, 6.37 ± 0.5Å, 6.02 ± 0.5Å, 5.61 ± 0.5Å, 5.37 ± 0.5Å, 5.18 ± 0.5Å, 4.95 ± 0.5Å, 4.82 ± 0.5Å, 4.69 ± 0.5Å, 4.53 ± 0.5Å, 4.35 ± 0.5Å, 4.23 ± 0.5Å, 4.07 ± 0.5Å, 3.88 ± 0.5Å, 3.82 ± 0.5Å, 3.77 ± 0.5Å, 3.71 ± 0.5Å, 3.57 ± 0.5Å, and 3.37 ± 0.5Å (d, 2d.p.).

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} Piperazin-1-yl] ethyl-1-one L-(+)-lactate C-type polymorphs are characterized by their interplanar spacing (d) values: 11.94 ± 0.2Å, 11.19 ± 0.2Å, 10.65 ± 0.2Å, 10.16 ± 0.2Å, 9.82 ± 0.2Å, 8.50 ± 0.2Å, 7.90 ± 0.2Å, 7.62 ± 0.2Å, 7.19 ± 0.2Å, 6.75 ± 0.2Å, 6.37 ± 0.2Å, 6.02 ± 0.2Å, 5.61 ± 0.2Å, 5.37 ± 0.2Å, 5.18 ± 0.2Å, 4.95 ± 0.2Å, 4.82 ± 0.2Å, 4.69 ± 0.2Å, 4.53 ± 0.2Å, 4.35 ± 0.2Å, 4.23 ± 0.2Å, 4.07 ± 0.2Å, 3.88 ± 0.2Å, 3.82 ± 0.2Å, 3.77 ± 0.2Å, 3.71 ± 0.2Å, 3.57 ± 0.2Å, and 3.37 ± 0.2Å (d, 2d.p.).

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} Piperazin-1-yl] ethyl-1-one L-(+)-lactate C-type polymorphs are characterized by their interplanar spacing (d) values: 11.94 ± 0.1Å, 11.19 ± 0.1Å, 10.65 ± 0.1Å, 10.16 ± 0.1Å, 9.82 ± 0.1Å, 8.50 ± 0.1Å, 7.90 ± 0.1Å, 7.62 ± 0.1Å, 7.19 ± 0.1Å, 6.75 ± 0.1Å, 6.37 ± 0.1Å, 6.02 ± 0.1Å, 5.61 ± 0.1Å, 5.37 ± 0.1Å, 5.18 ± 0.1Å, 4.95 ± 0.1Å, 4.82 ± 0.1Å, 4.69 ± 0.1Å, 4.53 ± 0.1Å, 4.35 ± 0.1Å, 4.23 ± 0.1Å, 4.07 ± 0.1Å, 3.88 ± 0.1Å, 3.82 ± 0.1Å, 3.77 ± 0.1Å, 3.71 ± 0.1Å, 3.57 ± 0.1Å, and 3.37 ± 0.1Å (d, 2d.p.).

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} The piperazine-1-yl] ethyl-1-one L-(+)-lactate C-type polymorph is characterized by its interplanar spacing (d) values: 11.94Å, 11.19Å, 10.65Å, 10.16Å, 9.82Å, 8.50Å, 7.90Å, 7.62Å, 7.19Å, 6.75Å, 6.37Å, 6.02Å, 5.61Å, 5.37Å, 5.18Å, 4.95Å, 4.82Å, 4.69Å, 4.53Å, 4.35Å, 4.23Å , 4.07Å, 3.88Å, 3.82Å, 3.77Å, 3.71Å, 3.57Å and 3.37Å (d, 2d.p.).

In yet another embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [ 3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} piperazine The C-type polymorph of L-(+)-lactate is characterized by its DSC peak temperature of 174.37ºC ± 0.5ºC (for example, 174.37ºC ± 0.2ºC, especially 174.37ºC ± 0.1ºC, more specifically 174.37ºC).

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} Piperazin-1-yl] ethyl-1-one L-(+)-lactate C-type polymorphs are characterized by a DSC onset temperature of 171.6ºC ± 0.5ºC (endothermic, sharp peak) (eg 171.6ºC ± 0.2 ºC, especially 171.6ºC ± 0.1ºC, and more specifically 171.6ºC).

In still another specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} The characteristics of the DSC temperature chart of the C-type polymorph of piperazin-1-yl] ethyl-1-one L-(+)-lactate are shown in FIG. 17 as 1.

In a specific embodiment, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3 , 2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} piperazine -1-yl] acet-1-one lactate (eg L-(+)-lactate)), which is crystalline and characterized by having one or more (in any combination) or all of the following parameters : (A) ¹ H NMR spectrum shown in Figure 15; and / or (b) XRPD pattern with peaks at: 7.4 ± 0.5 °, 7.9 ± 0.5 °, 8.3 ± 0.5 °, 8.7 ± 0.5 °, 9.0 ± 0.5 °, 10.4 ± 0.5 °, 11.2 ± 0.5 °, 11.6 ± 0.5 °, 12.3 ± 0.5 °, 13.1 ± 0.5 °, 13.9 ± 0.5 °, 14.7 ± 0.5 °, 15.8 ± 0.5 °, 16.5 ± 0.5 °, 17.1 ± 0.5 °, 17.9 ± 0.5 °, 18.4 ± 0.5 °, 18.9 ± 0.5 °, 19.6 ± 0.5 °, 20.4 ± 0.5 °, 21.0 ± 0.5 °, 21.8 ± 0.5 °, 22.9 ± 0.5 °, 23.3 ± 0.5 °, 23.6 ± 0.5 °, 24.0 ± 0.5 °, 24.9 ± 0.5 °, and 26.4 ± 0.5 ° (2θ, 1d.p); and / or (c) an XRPD pattern substantially as shown by mark 1 in FIG. 16; and / or (d) A peak having the same diffraction angle (2θ) as that of the XRPD pattern labeled 1 in FIG. 16, and selective, Where these peaks have the same relative intensity as the peak labeled 1 in FIG. 16; and / or (e) have the diffraction angle (2θ) and intensity of the main peak as shown in the XRPD pattern labeled 1 in FIG. 16; and / Or (f) the main peaks measured with XRPD are 8.7 ± 0.5º, 17.1 ± 0.5 °, 17.9 ± 0.5 ° and 18.9 ± 0.5 ° (2θ, 1d.p); and / or (g) the interplanar spacing ( d) Values: 11.94 ± 0.5 Å, 11.19 ± 0.5 Å, 10.65 ± 0.5 Å, 10.16 ± 0.5 Å, 9.82 ± 0.5 Å, 8.50 ± 0.5 Å, 7.90 ± 0.5 Å, 7.62 ± 0.5 Å, 7.19 ± 0.5 Å, 6.75 ± 0.5 Å, 6.37 ± 0.5 Å, 6.02 ± 0.5 Å, 5.61 ± 0.5 Å, 5.37 ± 0.5 Å, 5.18 ± 0.5 Å, 4.95 ± 0.5 Å, 4.82 ± 0.5 Å, 4.69 ± 0.5 Å, 4.53 ± 0.5 Å, 4.35 ± 0.5 Å, 4.23 ± 0.5 Å, 4.07 ± 0.5 Å, 3.88 ± 0.5 Å, 3.82 ± 0.5 Å, 3.77 ± 0.5 Å, 3.71 ± 0.5 Å, 3.57 ± 0.5 Å, and 3.37 ± 0.5 Å (d, 2d.p .); And / or (h) DSC peak temperature is 174.37ºC ± 0.5ºC (eg 174.37ºC ± 0.2ºC, especially 174.37ºC ± 0.1ºC, more specifically 174.37ºC); and / or (i) DSC onset The temperature is 171.6ºC ± 0.5ºC (endothermic, sharp peak) (such as 171. 6ºC ± 0.2ºC, especially 171.6ºC ± 0.1ºC, and more specifically 171.6ºC); and / or (j) DSC temperature chart as shown in Fig. 17 mark 1.

In particular, 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b ] Pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} piperazin-1-yl The polymorph form C of aceto-1-one L-(+)-lactate provides advantages regarding stability and crystallinity.

Complex

Chemical formula (I) also includes within its scope complexes of these compounds, where the complexes are, for example, inclusion complexes or clathrates with compounds, such as cyclodextrins, or metal complexes . Inclusion compounds, clathrates and metal complexes can be formed by those skilled in the art by well-known methods.

Prodrug

Formula (I) also includes any prodrug of a compound of formula (I). "Prodrug" means, for example, any compound that is converted into a biologically active compound of formula (I) in vivo.

For example, several prodrugs are esters of the active compound (eg, physiologically acceptable metabolizable labile esters). During metabolism, the ester group (-C (= O) OR) can be cleaved to form an active drug. Such esters can be formed, for example, by esterifying any of the carboxylic acid groups (-C (= O) OH) in the parent compound, if appropriate, before any other reactive group present in the parent compound. Protect, then deprotect if necessary.

Examples of such metabolizable labile esters include compounds of formula -C (= O) OR, where R is: C _1-7 alkyl (e.g., -Me, -Et, -nPr, -iPr, -nBu, -sBu, -iBu, -tBu); C _1-7 amine alkyl (e.g., amine ethyl; 2- (N, N-diethylamino) ethyl; 2- (4-morpholine) ethyl ); And ethoxy-C _1-7 alkyl (eg, ethoxymethyl; ethoxyethyl; trimethyl ethoxymethyl (pivaloyloxymethyl); acetoxymethyl (acetoxymethyl) ; 1-Ethyloxyethyl; 1- (1-methoxy-1-methyl) ethyl-carbonyloxyethyl (1- (1-methoxy-1-methyl) ethyl-carbonxyloxyethyl); 1 -(Benzyloxy) ethyl (1- (benzoyloxy) ethyl); isopropoxy-carbonyloxymethyl (1-propoxy-carbonyloxymethyl); -isopropoxy-carbonyloxyethyl); cyclohexyl-carbonyloxymethyl; 1-cyclohexyl-carbonyloxyethyl; cyclohexyloxy -carbonyloxymethyl); 1-cyclohexyloxy-carbonyloxyethyl; (4-tetrahydropyranyloxy) carbonyl (4-tetrahydropyranyloxy) carbonyloxymethyl; 1- (4-tetrahydropyranyloxy) carbonyloxyethyl; (4-tetrahydropyranyloxy) carbonyloxyethyl; ()-(4-tetrahydropyranyl) carbonyloxymethyl); and 1- (4-tetrahydropyranyl) carbonyloxyethyl) (1- (4-tetrahydropyranyl) carbonyloxyethyl).

In addition, several prodrugs can be activated by enzymes to obtain an active compound, or a compound can produce an active compound upon further chemical reactions (for example, antigen-directed enzyme prodrug therapy (ADEPT), gene-directed enzyme prodrug therapy (GDEPT), and ligand-directed enzyme prodrug therapy (LIDEPT), etc.). For example, the prodrug may be a carbohydrate derivative or other glucose conjugate, or may be an amino acid derivative. In one embodiment, the chemical formula (I) does not include the prodrug of the compound of the formula (I) in this range.

Advantages of the compounds of the invention

Compounds of formula (I) have several advantages over prior art compounds.

The compounds of the invention have particular advantages in one or more of the following aspects: (i) excellent selectivity for IKr (hERG) cardiac ion channels; (ii) excellent metabolic stability; (iii) lower liability for P450 inhibition (inhibitory liability); (iv) excellent oral bioavailability; and / or (v) excellent in vivo drug efficacy.

Excellent selectivity for IKr (hERG) cardiac ion channels

In the late 1990s, some US Food and Drug Administration-approved drugs were found to be dead and involved in cardiac abnormalities, and must be discontinued in the United States. It was then discovered that the side effect of these drugs was to block the hERG channel of the heart cells and lead to the development of arrhythmia. The hERG channel is a member of the potassium ion channel family, and the first member of the potassium ion channel family was identified in the mutant Drosophila melanogaster fruitfly in the late 1980s (see Jan, LY and Jan, YN (1990 ). A Superfamily of Ion Channels. Nature, 345 (6277): 672). The biophysical properties of the hERG potassium channel are described in Sanguinetti, MC, Jiang, C., Curran, ME, and Keating, MT (1995). A Mechanistic Link Between an Inherited and an Acquired Cardiac Arrhythmia: HERG encodes the Ikr potassium channel. Cell, 81: 299-307, and Trudeau, MC, Warmke, JW, Ganetzky, B., and Robertson, GA (1995). HERG, a Human Inward Rectifier in the Voltage-Gated Potassium Channel Family. Science, 269: 92 -95. Therefore, elimination of hERG blocking activity remains an important consideration in the development of any novel drug.

Many compounds of formula (I) have been found to have reduced hERG activity and / or a good separation between IAP activity and hERG activity (large "treatment window"). One method for measuring hERG activity is patch-clamped electrophysiology. Other methods of functional hERG activity measurement include hERG binding assays, which can use commercially available membranes isolated from cells stably expressing hERG channels or commercially available cell lines expressing hERG channels.

There are many compounds of formula (I) with improved Cardiac Safety Index (CSI) [CSI = hERG IC 50 / Cmax (unbound)] (Shultz et al, J. Med. Chem., 2011; Redfern et al , Cardiovasc. Res., 2003). This may be due to an increase in hERG IC 50 or a decrease in Cmax required for performance (due to better IAP efficacy and / or PK).

Certain compounds of formula (I) have reduced hERG ion channel blocking activity. The compound with specific formula (I) has an average IC ₅₀ value against hERG that is 30 times, or 40 times, or 50 times greater than the IC ₅₀ value of the compound in the cell proliferation assay. The average IC ₅₀ value against hERG possessed by the compound of the specific formula (I) is greater than 10 μM, more particularly greater than 20 μM, and more preferably greater than 30 μM. The average IC against several compounds of the present invention hERG _IC50 values greater than 40 uM lines, to indicate or display% inhibition at a concentration of 10, 30, ₅₀ or 300 uM of the IC. Several compounds of the invention have a higher average CSI than the minimum recommended value (30 times).

It can be seen from the data in Table 1 herein that the compounds of Examples 1 to 34 all have lower hERG reliability than the compound example 259 of WO 2012/143726 (the same is true for 262 and 263). In particular, the compounds of Examples 1 to 2, 11, and 34 of the present invention show an IC ₅₀ against hERG of ≥40 µM, whereas the compound example 259 of WO 2012/143726 (as well as 262 and 263) shows the hERG 42% inhibition. Therefore, the excellent selectivity for hERG is a key advantage of the compounds of the present invention over the IAP antagonist compounds disclosed in the prior art (especially those disclosed in WO 2012/143726).

Excellent metabolic stability

Compounds of formula (I) may have favorable ADMET properties, for example, better metabolic stability (as measured by mouse liver microsomes), a better P450 profile, and / or favorable clearance (E.g., low clearance). These characteristics allow more drugs to reach the appropriate site of action in the systemic circulation to exert its therapeutic effect, which is an advantage. Increasing the drug concentration in the tumor to exert the drug effect may increase the efficacy and therefore reduce the dosage. As such, compounds of formula (I) will exhibit reduced dosage requirements and will be easier to formulate and administer. In addition, the compound can reduce the turnover of P450 (eg 3A4).

Lower P450 inhibition liability

Many compounds of formula (I) are advantageous with different sensitivities to P450 enzymes. For example, a specific compound of formula (I) has an IC ₅₀ value greater than 10 µM against each of the cytochrome P450 enzymes 1A2, 2C9, 2C19, 3A4, and 2D6 (especially 3A4). In addition, these compounds are not P450 inhibitors in particular.

Excellent oral bioavailability

The compounds of the present invention potentially have physicochemical properties suitable for oral exposure (oral exposure or AUC). In particular, compounds of formula (I) may exhibit improved oral bioavailability. Oral bioavailability can be defined as the ratio (F) of a compound's plasma exposure when administered by the oral route to the compound's plasma exposure when administered by the intravenous (i.v.) route, expressed as a percentage.

Compounds with an oral bioavailability (F value, F%) greater than 30%, and more particularly greater than 40%, are particularly advantageous for oral administration without the use of non-oral administration, or non-oral administration may also be used.

Excellent in vivo efficacy

The compounds of the present invention can have improved in vivo pharmacological effects on cancer cell lines and in vivo models due to their improved pharmacological effects against XIAP and / or cIAP.

Preparation method of compound of formula (I)

This section is like the other sections of this application. Unless otherwise indicated in the text, the formula (I) also includes all other subgroups and examples thereof as defined herein.

Compounds of formula (I) can be prepared according to synthetic methods known to those skilled in the art.

According to another aspect of the present invention, a method for preparing a compound of formula (I) as described above is provided, which comprises:

(a) (i) combining a compound of formula (II): (II) wherein R ⁵ , R ⁶ , U and X are as described in the compound of formula (I), and L ¹ represents a suitable leaving group, such as a halogen atom (eg, chlorine) and P ¹ Represents hydrogen or a suitable protecting group, such as a third butoxycarbonyl group (tBoc), and reacts with a compound of formula (III): (III) or a selectively protected derivative thereof; wherein R ¹ and R ^{2 are} as previously described for the compound of formula (I), and then suitable for removing the P ¹ protecting group, and, if necessary, any other A protecting group, a deprotection reaction; or

(ii) the compound of formula (IV): (IV) wherein R ⁵ , R ⁶ , X and U are as previously described for the compound of formula (I), and L ² represents a suitable leaving group, such as a halogen (e.g., chlorine), and a compound of formula (V) reaction: (V) or a selectively protected derivative thereof; wherein R ¹ and R ^{2 are} as previously described for the compound of formula (I), and P ² represents hydrogen or a suitable protecting group such as a third butoxycarbonyl group ( tBoc), followed by a deprotection reaction suitable for removal of the P ² protecting group, and, if necessary, any other protecting group; and / or

(b) a deprotection reaction of a protected derivative of a compound of formula (I); and / or

(c) the interconversion of a compound of formula (I) or a protected derivative thereof into another compound of formula (I) or a protected derivative thereof; and

(d) Selectively forming a pharmaceutically acceptable salt of a compound of formula (I).

Step (a) (i) usually comprises the compound of formula (II) and the compound of formula (III), being selectively in the presence of a suitable additive such as potassium iodide and a suitable base such as potassium carbonate, in a suitable solvent such as acetonitrile. reaction. This step can be performed at ambient or elevated temperatures, such as 70ºC.

Steps (a) (ii) generally include the compound of formula (IV) and the compound of formula (V), being selectively in the presence of a suitable additive such as potassium iodide and a suitable base such as potassium carbonate in a suitable solvent such as acetonitrile. reaction.

Step (b) generally includes any suitable deprotection reaction, the conditions of which will depend on the nature of the protecting group. When the protecting group is tBoc, such a deprotection reaction will usually involve the use of a suitable acid in a suitable solvent. For example, the acid may suitably include trifluoroacetic acid or hydrogen chloride, and the solvent may suitably include dichloromethane ethyl acetate, 1,4-dioxane, methanol, or water. Optionally, a solvent mixture such as an aqueous methanol solution or ethyl acetate / 1,4-dioxane can be used.

It should be understood that when the protecting group is tBoc, when using a suitable acid as described above for deprotection, a compound of formula (I) which is a pharmaceutically acceptable salt can be produced, which can be isolated directly. Alternatively, the compound of formula (I) can be isolated in the form of a free base using methods known in the art, and then optionally converted into a pharmaceutically acceptable salt according to step (d).

Step (c) typically includes an interconversion step known to those skilled in the art. For example, in a compound of formula (I), a first substituent can be converted into a second, or another, substituent in a manner known to those skilled in the art. Technical Field This knows many known functional group interconversion reactions, the lead compound is converted to a compound of formula (the I), which can be found in Advanced Organic Chemistry by Jerry March, 4 th Edition, John Wiley & Sons, 1992. For example, possible metal-catalyzed functionalization reactions using organotin reagents (Stille reaction), Grignard Reagent, and reactions with nitrogen nucleophilic groups are described in 'Palladium Reagents and Catalysts' [Jiro Tsuji, Wiley, ISBN 0-470-85032-9] and Handbook of OrganoPalladium Chemistry for Organic Synthesis [Volume 1, Edited by Ei-ichi Negishi, Wiley, ISBN 0-471-31506-0].

Step (d) can be carried out by dissolving the compound of formula (I) in free base in a suitable solvent, treating with a chemical dose or excess of a pharmaceutically acceptable organic or non-acidic acid, and then using the obtained salt as a technique Separation is performed by methods known in the art (e.g., solvent evaporation or crystallization).

Where appropriate, one or more reactions known to those skilled in the art may be followed or preceded by the reactions of steps (a), (b), and (c), and performed in the appropriate order to define R ¹ , R ^The necessary substitutions are made on ² , R ⁵ , and R ⁶ to obtain other compounds of formula (I). The conditions for such reactions have been described in the literature, and non-limiting examples include: reactive functional group protection reactions; reactive functional group deprotection reactions; halogenation reactions; dehalogenation reactions; dealkylation reactions; amines, anilines, Alkylation and arylation of alcohols and phenols; Mitsunobu reaction on hydroxyl groups; Cycloaddition reactions on appropriate groups; Reduction reactions on nitro, ester, cyano, and aldehyde groups; Transition Metal-catalyzed coupling reaction; acetylation reaction; sulfonation reaction / sulfonyl group introduction reaction; saponification reaction / ester hydrolysis reaction; ester amidation reaction or transesterification reaction; carboxyl ester Reaction or halogenation reaction; halogen exchange reaction; nucleophilic substitution reaction using amine, thiol or alcohol; reductive amination reaction; oxime formation reaction on carbonyl group or hydroxylamine group S-oxidation reaction; N-oxidation reaction; and salification reaction.

Compounds of formula (II) can be prepared from compounds of formula (IV) according to the following scheme 1 (Scheme 1): Among them, X, U, R ⁵ , R ⁶ , L ¹ , L ² and P ^{1 are} as defined above.

Step (i) of Scheme (1) usually comprises making the compounds of formula (IV) and formula (VI), selectively in the presence of a suitable additive such as potassium iodide and a suitable base such as potassium carbonate in a suitable solvent (such as acetonitrile)中 反应。 In the reaction.

When L ¹ represents chlorine, step (ii) of Scheme 1 usually comprises combining a compound of formula (VII) with a reagent capable of converting a hydroxyl group into a good leaving group (such as methanesulfonium) in the presence of a base such as triethylamine. Chlorine) reaction.

The compound of formula (IV) can be prepared according to the following scheme 2, wherein X represents N, U represents carbon, and R ⁶ represents methylol: Scheme 2

Among them, L ³ , L ⁴ , L ⁵ and L ⁶ represent suitable leaving groups such as a halogen atom (ie, fluorine, bromine or chlorine) and R ⁵ and L ² are as described above.

When both L ³ and L ⁴ represent fluorine, step (i) of Scheme 2 generally comprises combining a compound of formula (VIII) with a base, such as sodium bis (trimethylsilyl) amide, in Tetrahydrofuran and isobutyronitrile are reacted in a suitable solvent such as toluene. An example of such a reaction is shown in Preparation Example 11 herein.

Step (ii) of Scheme 2 involves reacting with a suitable reducing agent, which typically comprises reacting a compound of formula (IX) with a borane-tetrahydrofuran complex in the presence of a suitable solvent such as tetrahydrofuran. An example of such a reaction is shown in Preparation Example 12 herein. Step (ii) of Scheme 2 may also generally include reacting a compound of formula (IX) with nickel (II) chloride hexahydrate, and then adding sodium borohydride. An example of such a reaction is shown in Preparation 12 herein, and another step option.

Step (iii) of Scheme 2 generally comprises cyclisation of a compound of formula (X), using a suitable base, such as potassium carbonate, and a suitable solvent, such as NMP. An example of such a reaction is shown in Preparation Example 13 herein.

Step (iv) of Scheme 2 generally comprises reacting a compound of formula (XI) with a compound of formula R ⁵ -M, wherein R ^{5 is} as described above, and M represents an organometallic residue, such that R ⁵ -M represents nucleophilicity. Organometallic reagents, such as organozinc halide. Step (iv) also typically involves the use of lithium bromide, a catalyst such as [1,3-bis (2,6-diisopropylphenyl) imidazol-2-ylidene] (3-chloropyridyl) palladium (II) dichloride ) ([1,3-bis (2,6-diisopropylphenyl) imidazol-2-ylidene] (3-chloropyridyl) palladium (II) dichloride), which is used in suitable solution systems such as tetrahydrofuran and NMP. An example of such a reaction is shown in Preparation 15 herein.

Step (v) of Scheme 2 usually comprises halogenation of a compound of formula (XII), for example, halogenation using N-bromosuccinimide in dimethylformamide. An example of such a reaction is shown in Preparation 16 herein.

Step (vi) of Scheme 2 involves lithiation and reaction with a suitable electrophile to introduce formamidine, and generally involves reacting a compound of formula (XIII) with MeLi in THF and then adding it to TBuLi in hexane and then dimethylformamide. An example of such a reaction is shown in Preparation Example 17 herein.

Step (vii) of Scheme 2 involves the reduction of methylamyl using a suitable reducing agent, and typically involves reacting a compound of formula (XIV) with sodium borohydride in methanol. An example of such a reaction is shown in Preparation Example 17 herein.

When L ² represents a halogen atom, such as chlorine, step (viii) of Scheme 2 usually comprises reacting a compound of formula (XV) with a haloacetyl halide such as chloroethenyl chloride in MeCN, and then adding it to Potassium carbonate in methanol. An example of such a reaction is shown in Preparation 18 herein. Another option is to convert the compound of formula (XIII) into a compound of formula (XV) by following a sequence similar to that described in Preparation Examples 25-29.

It should be understood that the compound of formula (XV), in which R ⁶ represents CH (OR ^x ) CH ₂ OR ^z , can be prepared in a manner similar to the above scheme 2 by changing steps from step (v) of scheme 2 . Examples of suitable reaction sequences are shown in Preparation Examples 38 to 42 herein.

Compounds in which X represents NR ³ , U represents carbon, and R ⁶ is = 0, can be synthesized by converting functional groups to each other through an appropriate intermediate of Scheme 2 or a protected derivative thereof, for example, Preparation Examples 22 to 24, 30 ~ 35 and 50.

It should be understood that the compound of formula (IV) in which R ⁵ represents unsubstituted n-butyl or in another case a substituted benzyl group, by changing the organometallic reagent used in step (iv) of Scheme 2, It can be prepared in a manner similar to Scheme 2. An example of such a reaction is shown in Preparation Examples 15A, 15B and 15C herein.

A compound in which X represents CR ⁴ , U represents nitrogen, and R ⁶ represents a pendant oxygen group can be synthesized using sequences similar to those described in Preparation Examples 43 to 49 and 51 to 58.

The compound of formula (V), or a selectively protected derivative thereof, can be prepared according to the following scheme 3: Scheme 3

Among them, R ¹ , R ² and P ^{2 are} as defined above for the compound of formula (V), L ⁷ represents a suitable leaving group, such as a halogen atom (for example, chlorine), and P ³ represents a suitable protecting group, such as Benzyl.

When P ³ represents benzyl, step (i) of Scheme 3 usually includes bringing the compound of formula (XVI) and benzaldehyde in a suitable reducing agent such as sodium triacetoxyborohydride and 1,2- Reaction in the presence of dichloroethane. An example of such a reaction is shown in Preparation Example 5 herein.

When L ⁷ represents chlorine, step (ii) of Scheme 3 generally includes reacting a compound of formula (XVII) with methanesulfonyl chloride in the presence of triethylamine and dichloromethane. An example of such a reaction is shown in Preparation Example 6 herein.

Step (iii) of Scheme 3 generally comprises reacting the compounds of formula (XVIII) and (XIX) in a suitable solvent (such as acetonitrile) in the presence of a base (such as potassium carbonate) and an additive (such as potassium iodide). An example of such a reaction is shown in Preparation Example 7 herein.

Step (iv) of Scheme 3 usually includes a deprotection reaction. For example, when P ³ represents benzyl, step (iv) usually comprises a hydrogenation reaction of a compound of formula (XX), which is in the presence of a suitable catalyst (such as palladium on carbon) in a suitable solvent system (Such as ethanol or a mixture of acetic acid and ethanol. An example of such a reaction is shown in Preparation Example 8 herein.

Alternatively, a compound of formula (I) can be obtained by making a compound of formula (XXI) or a selectively protected derivative thereof: (XXI) wherein R ¹ and R ^{2 are} as defined in the aforementioned compound of formula (I), and P ² represents a suitable protecting group, such as a third butoxycarbonyl group (tBoc), which is synthesized by reacting with a compound of formula (XXII) : (XXII) wherein X, U, R ⁵ and R ^{6 are} as described above, and then a deprotection reaction suitable for removing the protecting group P ² and any other protecting groups is performed.

An example of a suitable compound of formula (XXII) includes a compound of formula (XV) as described above.

This reaction usually involves combining a compound of formula (XXI) with a compound of formula (XXII), such as a compound of formula (XV), in a suitable solution and at a suitable temperature (for example, ambient temperature), in which the compound of formula (XXI) has Carboxyl groups react in the presence of suitable bases and reagents. A suitable solvent (such as dichloromethane) should be inert to the reagents used. Examples of suitable bases are triethylamine and N, N-diisopropylethylamine (DIPEA). Examples of suitable activators are bromo-tris-pyrrolidino-phosphonium hexofluorophosphate (PyBrop), O-benzotriazole-N, N, N ', N'-tetramethyl O-benzotriazole-N, N, N ', N'-tetramethyl-uronium-hexafluoro-phosphate (HBTU), 1,1'-carbonyldiimidazole ), 1-ethyl-3- (3'-dimethylaminopropyl) -carbodiimide hydrochloride (EDC), And 2- (7-nitro-1H-benzotriazol-1-yl) -1,1,3,3-tetramethylurea cation hexafluorophosphate (2- (7-aza-1H-benzotriazole-1 -yl) -1,1,3,3-tetramethyluronium hexafluorophosphate (HATU). This method can be selectively used in the presence of a catalytic amount or a stoichiometric suitable coactivation reagent such as 1-hydroxybenzotriazole (HOBt) or 1-hydroxyazabenzotriazole (HOAt). Carry on.

The compound of formula (XXI) or a selectively protected derivative thereof can be prepared from a compound of formula (V) or a selectively protected derivative thereof as described above in a manner known in the art, for example, an ester with monohaloacetic acid ( (E.g. benzyl bromoacetate) in the presence of a suitable base (e.g. potassium carbonate) in a suitable solvent (e.g. acetonitrile); and then an ester hydrolysis reaction (or in the case of phenylmethyl ester) Selectively perform a hydrogenolysis reaction). The compound of formula (I) can be prepared by following a sequence similar to that described in Preparations 1 to 5.

Compounds of formula (XXII) can be prepared using sequences similar to those in Scheme 2 or the following preparations: 38-42; 22-24, 30-35, and 50; or 43-49 and 51-58.

It should be understood that specific compounds such as formulas (I), (II), (III), (V), (VI), (VII), (XVI), (XVII), (XVIII), (XIX), (XX ), (XXI) and (XXII) compounds can exist in different forms of non-mirromeric isomers and / or mirror-isomeric isomers, and their preparation methods can be synthesized as precursors of pure mirror-isomers.

Alternatively, racemic precursors can be used, and mixtures of non-diastereoisomers produced in these methods can be separated by methods known to those skilled in the art, such as using non-palm or palm Preparative preparative chromatography or resolution using non-mirromeric isomers: for example, crystals that form a salt with an enantiomerically pure acid (such as L-tartaric acid); or a pure mirror image Heterogeneous palm adjuvants are covalently bonded to the compound to form non-mirror isomerized derivatives, which are then separated using conventional methods such as chiral chromatography, and the aforementioned covalent bonds are then cleaved It is cut to produce the appropriate enantiomerically pure product, and the enantiomer separation is achieved.

Desired intermediates, such as compounds of formulae (III), (VI), (VIII), R ⁵ -M, (XVI) and (XIX), are commercially available, are known in the literature, and are similar to those described in the literature Prepared by methods, or by methods similar to the experimental steps described in the following examples. Other compounds can be prepared by converting functional groups of R ¹ , R ² , R ⁵ and R ⁶ groups using methods well known in the art.

In yet another embodiment, the present invention provides a novel intermediate. In a specific embodiment, the present invention provides novel intermediates having formula (II) or (IV) or (V) or (VII) or (XX). In another embodiment, the present invention provides a novel intermediate having the formula (XXI) or (XXII).

Protection group

In many of the reactions described above, it may be necessary to protect one or more groups to prevent undesired reactions on the molecule from occurring. For examples of protecting groups, and methods for protecting and deprotecting functional groups, see Protective Groups in Organic Synthesis (T. Green and P. Wuts; 3rd Edition; John Wiley and Sons, 1999).

In particular, the R ¹ and R ² groups can be synthesized in a protected form, and removal of the protecting group can produce a compound of formula (I).

The hydroxyl group can be protected, for example, as an ether (-OR) or an ester (-OC (= O) R), for example, it can form: a third butyl ether; a tetrahydropyran (THP) ether; benzyl , Benzhydryl (diphenylmethyl), or trityl (triphenylmethyl) ether; trimethylsilyl or third butyldimethylsilyl ether; or acetamyl ester (-OC ( = O) CH ₃ ).

Aldehydes or keto groups can be protected, for example, acetal (R-CH (OR) ₂ ) or ketal (R ₂ C (OR) ₂ ), respectively, where the carbonyl group (> C = O) is treated with, for example, a primary alcohol. In the presence of an acid, a large excess of water is used to carry out the hydrolysis reaction, which can conveniently regenerate the aldehyde or ketone group.

Amine group may be protected, for example, may be formed Amides (-NRCO-R) or carbamate (carbamate) (-NRCO-OR) , for example, may be formed: A Amides (-NHCOCH ₃₎ ; Benzyl carbamate (-NHCO-OCH ₂ C ₆ H ₅ , -NH-Cbz or NH-Z); third butyl carbamate (-NHCO-OC (CH ₃ ) ₃ , -NH -Boc); 2-diphenyl-2-propylcarbamate (-NHCO-OC (CH ₃ ) ₂ C ₆ H ₄ C ₆ H ₅ , -NH-Bpoc); 9-fluorenylmethylcarbamate 9-fluorenylmethyl carbamate (-NH-Fmoc); 6-nitroveratryl carbamate (-NH-Nvoc); 2-trimethylsilyl ethyl carbamate ( -NH-Teoc); 2,2,2-trichloroethyl carbamate (-NH-Troc); allyl carbamate (-NH-Alloc); or 2 (-phenylfluorenyl) Ethyl carbamate (-NH-Psec).

For example, if the compound of formula (II) contains an amine group, when additional functional groups are to be introduced, the amine group can be protected by the aforementioned protecting group, and a special protecting group is the third butoxycarbonyl group ( Boc). When the amine group does not require subsequent modification reaction, a protecting group can be attached to give the compound N-protected form through the reaction sequence, and then the general method (for example, acid treatment in the Boc group can be used) for deprotection, and A compound of formula (I) is obtained.

Other protective groups of amine groups, such as cyclic amines and heterocyclic NH groups, include tosyl and mesyl groups, benzyl (eg, p-methoxybenzyl (PMB)) groups And tetrahydropyran (THP) groups.

The carboxylic acid group may be protected as an ester, for example, it may form: a C _1-7 alkyl ester (e.g., methyl ester; third butyl ester); a C _1-7 haloalkyl ester (e.g., C _1-7 trihaloalkyl ester); tri C _1-7 alkylsilyl-C _1-7 alkyl ester; or C _5-20 aryl-C _1-7 alkyl ester (eg, benzyl Esters; nitrobenzyl esters; p-methoxybenzyl esters). Thiols can also be protected in the manner of, for example, thioethers (-SR). For example, they can be formed: benzyl sulfide; acetamidomethyl ether (-S-CH ₂ NHC ( = O) CH ₃ ).

Isolation and purification of compounds of the present invention

The compounds of the present invention can be isolated and purified by general techniques known to those skilled in the art, and examples of this method include chromatography techniques such as column chromatography (e.g., flash chromatography) and HPLC. One particularly useful technique for compound purification is preparative liquid chromatography, which uses mass spectrometry to detect compounds flowing out of a chromatography column.

Preparative LC-MS is a common and effective method for purifying small molecule organic compounds (such as the compounds described herein). The methods of liquid chromatography (LC) and mass spectrometry (MS) can be changed to provide better separation of crude products and improve sample detection of MS. The optimization conditions of the preparative gradient LC method include the use of different columns, volatile eluents and modifiers, and the eluent gradient. Compounds can be purified by these methods using optimized preparative LC-MS methods known in the art. Such methods are Rosentreter U, Huber U .; Optimal fraction collecting in preparative LC / MS; J Comb Chem .; 2004; 6 (2), 159-64 and Leister W, Strauss K, Wisnoski D, Zhao Z, Lindsley C., Development of a custom high-throughput preparative liquid chromatography / mass spectrometer platform for the preparative purification and analytical analysis of compound libraries; J Comb Chem .; 2003; 5 (3); 322-9. An example of a system for purifying compounds by preparative LC-MS is as described in the Examples section of the present invention below (the title "Quality-Directed Purification LC-MS System").

The recrystallization method of the compound of formula (I) and its salt can be performed using methods known to those skilled in the art, see, for example, P. Heinrich Stahl (Editor), Camille G. Wermuth (Editor), ISBN: 3-90639- 026-8, Handbook of Pharmaceutical Salts: Properties, Selection, and Use, Chapter 8, Publisher Wiley-VCH. Products obtained from organic reactions are rarely pure if they are separated directly from the reaction mixture. When the compound (or its salt) is a solid, it can be purified and / or crystallized by a recrystallization method from an appropriate solvent. A good recrystallization solvent must be able to dissolve an appropriate amount of the substance to be purified at an elevated temperature, but only a small amount of the substance can be dissolved at a low temperature. It must also dissolve impurities at low temperatures, or incompletely dissolve impurities. Finally, the solvent should be easily removed from the purified product. This usually means that the solvent has a relatively low boiling point and that those skilled in the art will understand the recrystallization solvents used for a particular substance; or if this information is not available, multiple solvents must be tested. In order to obtain a good yield of purified material, a minimum amount of hot solvent is used to dissolve all impurities. In fact, 3 to 5% more solvent is used than necessary to make the solution unsaturated. If the impure compound contains impurities that cannot be dissolved in the solvent, it can be subsequently removed by filtration, and then the solution is crystallized. In addition, if the impure compound contains a small amount of non-compound colored substances, a small amount of a decolorizing agent (such as activated carbon) can be added to the hot solution, and the solution can be crystallized after filtering. Crystallization usually occurs spontaneously when the solution is cooled. However, if it does not proceed spontaneously, the solution can be cooled to below room temperature or a single crystal (seed) of pure substance can be added to promote the crystallization. Recrystallization can also be performed and / or anti-solvent or co-solvent can be used to optimize the yield. In this case, the compound is dissolved in a suitable solvent at elevated temperature, and another solvent is added after filtration to help recrystallization, wherein the desired compound has a low solubility in this additional solvent. Thereafter, the obtained crystals can generally be separated using a vacuum filtration method, and then washed and dried in an oven or a drying method, for example.

Other examples of purification methods include: sublimation, which includes a step of heating in a vacuum environment (for example, using a cold finger condenser); and crystallization from a molten state (Crystallization Technology Handbook 2nd Edition, edited by A Mersmann, 2001).

Biological effects (BIOLOGICAL EFFECTS)

The compounds, subgroups, and examples thereof of the present invention act as antagonists of apoptosis inhibitory proteins (IAPs) and are effective in preventing or treating the disease states or abnormal physical conditions described herein. In addition, the compounds of the present invention and its subgroups are also effective in preventing or treating diseases or abnormal physical conditions mediated by IPA. Refers to the prevention or suppression or treatment of a disease state or abnormal physical condition such as cancer, including slowing or reducing the induction of cancer within its scope.

Thus, for example, the compounds of the invention will be useful in slowing or reducing the incidence of cancer.

The compounds of the present invention are useful in the treatment of adults. The compounds of the invention are also useful in the treatment of children.

More particularly, compounds of formula (I) and their subgroups are antagonists to IAP. For example, the compounds of the invention have an affinity for XIAP, cIAP1 and / or cIAP2, and in particular an IAP selected from XIAP and cIAP1.

Specific compounds are compounds that have an affinity for one or more IAPs selected from XIAP, cIAP1, and cIAP2. A specific compound of the present invention is a compound having an IC ₅₀ value of less than 0.1 μM.

Antagonist Compounds of formula (I) are capable of binding to IAP and exhibiting efficacy against IAP. In a specific embodiment, the compound of formula (I) of the antagonist can exhibit selectivity to one or more IAP members in the IAP family member. Compared with binding and / or exhibiting affinity with other members of the IAP family, the antagonist formula (I) The compounds are more likely to bind to XIAP and / or cIAP and / or show affinity for XIAP and / or cIAP.

In addition, many compounds of the present invention are selective for XIAP over cIAP, or vice versa, that is, they are selective for cIAP over XIAP (especially cIAP1), and a specific embodiment of the present invention illustrates these compounds. In particular, compared to other IAP family members, the compounds of the present invention can exhibit at least ten times more affinity for one or more, especially XIAP, cIAP1, and / or cIAP2 members of the IAP family. This can be determined by the methods described herein. In yet another embodiment, the compounds of the present invention may have comparable affinities to XIAP, cIAP1, and / or cIAP2, and in particular, have comparable affinities to XIAP and cIAP1 (ie, differences in affinity are less than 10-fold).

It can be particularly advantageous for the activity of XIAP and cIAP1. To antagonize XIAP and cIAP1 with the same potency, it should be able to initiate the apoptotic response by activating caspase-8, and to switch from the pro-survival NF-κB message to the direction of apoptosis; and the possibility of XIAP Antagonism ensures that the apoptotic effect is reached before any inherent impedance mechanism used to block this process is upregulated. When cIAP1 is depleted through self-ubiquitination and proteasome degradation, NF-κB signaling will temporarily rise and regulate, which responds to the expression of TNF-α in sensitive cell lines, and also responds to cIAP2 and c-FLIP. Anti-Apoptotic Factors Rise Regulated Responses. Therefore, an effective XIAP antagonistic mechanism is needed to effector caspase activation and cell death, rather than allowing cIAP2-mediated accumulation of resistance. It is generally believed that the toxic response caused by the administration of these compounds in the body is due to the temporary induction of NF-κB signaling, resulting in the up-regulation of pro-inflammatory cytokines. This response is mediated by the cIAP1 / 2 antagonistic mechanism alone . Therefore, this dual effect should be able to form a therapeutic window before a dose-limiting toxic response occurs.

The function of IAP in controlling programmed cell death is also related to a variety of diseases, including disorders related to cell aggregation (e.g., cancer, autoimmune disorders, inflammatory responses and cardiovascular restenosis), excessive Deregulation of cell loss due to apoptosis (e.g., stroke, heart failure, neurodegeneration such as Alzheimer's disease, Parkinson's disease, Huntington's disease, amyotrophic lateral sclerosis, AIDS, ischemia (Stroke, myocardial infarction) and osteoporosis, or treatment of autoimmune diseases such as multiple sclerosis (MS).

Therefore, it is also envisaged that the compounds of the present invention can be used to treat other abnormal physical conditions such as inflammation (such as arthritis including rheumatoid arthritis), hepatitis, ulcerative colitis, gastritis, autoimmune disease, cardiovascular restenosis , Stroke, heart failure, neurodegenerative abnormal physical conditions (e.g., Alzheimer's disease, Parkinson's disease, Huntington's disease, tonic muscular dystrophy, and amyotrophic lateral sclerosis), AIDS, ischemia (e.g., traumatic brain injury, spinal injury, cerebral ischemia, cerebral ischemia / perfusion (I / R) injury, acute and chronic CNS injury ischemia, stroke or myocardial infarction), musculoskeletal Systems such as degenerative diseases of osteoporosis, autoimmune diseases (e.g., multiple sclerosis (MS) and type 1 diabetes), and eye diseases (e.g., retinal degenerative diseases due to programmed cell death control disorders) . In a specific embodiment, the compounds of the present invention can be used to treat viral infections such as herpes virus, pox virus, Epstein-Barr virus, Sindbis virus, adenovirus, HIV, HPV, hepatitis such as hepatitis B virus (HBV) or hepatitis C virus. Hepatitis virus (HCV) and HCMV or mycobacterial infections such as tuberculosis (TB).

Due to the affinity of the compounds of the present invention for IAP, it will be useful as a means to control programmed cell death. The compounds are therefore expected to be useful in the treatment or prevention of proliferative disorders such as cancer. In addition, the compounds of the present invention can be used for the treatment of diseases such as disorders related to cell aggregation or disorders caused by excessive apoptosis.

Examples of cancers (and their benign tissues) that can be treated (or inhibited) include, but are not limited to, epithelial origin tumors (various forms of adenomas and malignancies, including adenocarcinoma, squamous cell carcinoma, transitional Cell carcinoma or other malignant tumors), such as the bladder and urethra, breast, gastrointestinal tract (including esophagus, stomach, small intestine, large intestine, rectum and anus), liver (liver cancer), gallbladder and bile duct system, exocrine pancreas, kidney, lung ( For example, adenocarcinoma, small cell lung cancer, non-small cell lung cancer, bronchioalveolar carcinomas and mesothelioma), head and neck (e.g. tongue, mouth, throat, pharynx, nasopharynx, tonsil, salivary glands, nasal cavity And sinuses), ovaries, fallopian tubes, peritoneum, vagina, vulva, penis, cervix, myometrium, endometrium, thyroid (e.g., follicular thyroid cancer), adrenal gland, prostate, skin and skin components (e.g., Melanoma, basal cell tumor, squamous cell tumor, keratoacanthoma, dysplastic nevus) and other malignant tumors; hematological malignancies (ie, leukemia, lymphoma), and hematological malignancies Precancerous blood diseases of the lymphatic system and borderline malignancies (eg, acute lymphocytic leukemia [ALL], chronic lymphocytic leukemia [CLL], B-cell lymphomas such as diffuse large B-cell lymphoma [DLBCL], follicles Lymphoma, Burkitt's lymphoma, mantle cell lymphoma, T-cell lymphoma and leukemia, natural killer cell [NK] lymphoma, Hodgkin's lymphoma, hair cell leukemia, monoclonal gammopathy of unknown significance , Plasmacytoma, multiple myeloma, and lymphoproliferative disorders after transplantation, and hematological malignancies and disorders of the bone marrow system (for example, acute myeloid leukemia [AML], chronic myeloid leukemia [CML], chronic bone marrow mononuclear Globular leukemia [CMML], hypereosinophilic syndrome, myeloproliferative disorders such as polycythaemia vera, primary thrombocytosis and primary myelofibrosis, Malignant tumors such as myelodysplastic syndromes, myelodysplastic syndromes, and promyelocytic leukemias); mesenchymal tumors , For example, soft tissue, bone or cartilage sarcomas, such as osteosarcoma, fibromas, chondrosarcoma, rhabdomyosarcoma, leiomyosarcoma, liposarcoma, angiosarcoma, Kaposi 'sarcoma, Ewing sarcoma, synovial cell tumor, epithelial sarcoma Gastrointestinal stromal tumors, benign and malignant histiocytomas, and bulging dermal fibromas; tumors of the central or peripheral nervous system (e.g., astrocytoma, glioma or glioblastoma, meningiomas, ependyma Tumors, pineal tumors, and Schwann cell tumors); endocrine tumors (for example, pituitary tumors, adrenal tumors, islet cell tumors, parathyroid tumors, carcinoid tumors, and thyroid medullary tumors); eyes and eyes Accessory tumors (eg, retinoblastoma); germ cells and trophoblastomas (eg, teratomas, seminoma, malignant embryo tumors, hydatidiform moles, and chorionic carcinoma); And children and embryo tumors (e.g., medulloblastoma, neuroblastoma, Wilms tumor, and primitive neuroectodermal tumors); or syndromes, congenital or volume Resulting in the case of proliferative disorders (e.g., xeroderma pigmentosum disease).

Cell growth is a tightly controlled effect. Cancer is an abnormal cell growth condition when cells replicate in an uncontrolled manner (increase in number), uncontrolled cell growth (larger), and / or due to apoptosis (programmed cell death), gangrene, or Cell death caused by detachment is reduced and cancer develops. In a specific embodiment, the abnormal cell growth is selected from the group consisting of: uncontrolled cell proliferation, excessive cell growth, or reduced programmed cell death. In particular, the condition or disease of abnormal cell growth is cancer. Therefore, in a pharmaceutical composition, use, or method for treating a disease or abnormal physical condition including abnormal cell growth (ie, uncontrolled and / or rapid cell growth) of the present invention, abnormal cells are included in a specific embodiment The growing disease or abnormal physical condition is cancer.

In a specific embodiment, the malignant blood disease is leukemia. In another specific embodiment, the malignant blood disease is lymphoma.

In a specific embodiment, the disease to be treated is leukemia, such as acute and chronic leukemia, acute myeloid leukemia (AML), and chronic lymphocytic leukemia (CLL). In a specific embodiment, the leukemia is refractory DLBCL.

In a specific embodiment, the lymphoma is MALT lymphoma. In a specific embodiment, the leukemia is AML.

In a specific embodiment, the malignant blood disease is multiple myeloma.

Many diseases are characterized by continuous and unregulated angiogenesis. Chronic proliferative diseases are generally highly related to angiogenesis, which may cause or maintain an inflammatory and / or proliferative state, or cause tissue damage due to invasive proliferation to blood vessels. Tumor growth and metastasis have been shown to be related to angiogenesis. Therefore, the compounds of the present invention can be used to prevent and disturb the initiation of tumor angiogenesis. In particular, the compounds of the invention are useful in the treatment of metastatic and metastatic cancer.

Metastatic or metastatic disease is the transfer of disease from one organ or part to another non-adjacent organ or part. Cancers treatable by the compounds of the present invention include primary tumors (i.e., tumor cells in the primary location), regionally invasive cancers (cancer cells pass through or infiltrate surrounding normal tissue in a localized area), and metastasis (Or secondary) tumors, that is, tumors that begin to form from malignant cells, where the malignant cell line progresses through the bloodstream (blood-type spread) or through the lymph or through the body cavity (spatial metastasis) to other areas and tissues of the body cycle.

Specific cancers include liver cancer, melanoma, esophageal cancer, kidney cancer, colon cancer, rectal cancer, lung cancer (eg, mesothelioma or lung adenocarcinoma), breast cancer, bladder cancer, gastric cancer, ovarian cancer, and prostate cancer.

Examples of specific cancers include kidney cancer, melanoma, colon cancer, lung cancer, breast cancer, ovarian cancer, and prostate cancer. In a specific embodiment, the cancer is selected from the group consisting of melanoma, colon cancer, breast cancer, and ovarian cancer. In a specific embodiment, the cancer is melanoma. In a specific embodiment, the cancer is inflammatory breast cancer.

In a specific embodiment, the cancer is lung cancer, such as mesothelioma, which includes malignant peritoneal mesothelioma or malignant pleural mesothelioma.

In a specific embodiment, the cancer is breast cancer, especially triple-ve breast cancer.

In a specific embodiment, the cancer is rectal cancer.

Yet another aspect of the present invention includes the use of a compound of the present invention to prevent or treat cancer in a patient, wherein the patient is selected from a subgroup of cancers with a highly inflammatory condition. Such cancers are also known as "inflammatory manifestations" and include cancers that increase cytokine signaling (eg, TNF). In a specific embodiment, the cancer is an inflammatory tumor. For example, the cancer is melanoma, colon cancer, breast cancer, and ovarian cancer, and particularly melanoma.

In a specific embodiment, the melanoma is a ras gene mutant melanoma.

Some cancer lines are resistant to specific medications. This cause may be related to the type of tumor (most commonly the epithelial malignant tumor itself is chemically resistant), or the resistance may arise spontaneously due to disease progression or treatment results. In this regard, examples of mesothelioma resistance include mesothelioma resistance to topoisomerase poisons, methylation agents, antitubulin agents, antifolates, platinum compounds, and radiation therapy, Especially anti-cisplatin mesothelioma. Similarly, examples of multiple myeloma include bortezomib-sensitive multiple myeloma or refractory multiple myeloma, and examples of chronic myeloid leukemia include imitanib-sensitive Chronic myelogenous leukemia and refractory chronic myelogenous leukemia.

The cancer may be sensitive to an antagonistic response to any one or more of IAP selected from XIAP, cIAP1, cIAP2, NAIP, ILP2, ML-IAP, survivin, and BRUCE, preferably XIAP, cIAP1, cIAP2, ML- IAP, and more preferably XIAP.

It is further envisaged that the compounds of the present invention, especially those having an affinity for IAP, may be particularly useful in the treatment or prevention of cancers associated with increased IAP or 11q22 amplification, and examples of cancers are as described in the prior art herein.

In a variety of cancers, an increase in the amount of IAP due to IAP overexpression is found, and an increase in the amount of IAP causes poor prognosis. In addition, cancers associated with 11q22 amplification are also sensitive to IAP antagonists. Increased IAP volume and 11q22 amplification can be identified using the techniques described herein. Regardless of whether a particular cancer is sensitive to IAP function, it can be detected by the method described in the paragraph "Diagnostic Methods" herein.

Yet another aspect of the present invention provides the use of a compound for the manufacture of a therapeutic agent for a disease or abnormal physical condition described herein, particularly cancer.

The compounds of the present invention can also be used to treat tumor growth, occurrence, chemical and radiotherapy resistance of cells sensitized to chemotherapy, and as anti-metastatic agents.

All types of anti-cancer interventions must increase the pressure on target tumor cells. In slowing down the harmful reactions caused by this stress, IAP is a direct anti-cancer drug and treatment. Therefore, the antagonist of IAP represents a chemotherapeutic agent with the ability to: (i) increase the sensitivity of harmful cells to anticancer drugs and / or treatments; (ii) alleviate or reduce the anticancer drugs and / or treatments Incidence of resistance; (iii) altering resistance to anticancer drugs and / or treatments; (iv) exhibiting anticancer drug and / or treatment activity; (V) delaying or preventing resistance to anticancer drugs and / or treatments Sexuality.

As a result of its affinity for IAP, compounds can be used in methods to control programmed cell death. Therefore, it is conceivable that the compounds of the present invention can be used to treat other abnormal physical conditions: such as inflammatory reactions caused by hepatitis, ulcerative colitis, and gastritis; such as Alzheimer's disease, Parkinson's disease, Huntington's disease, rigidity Muscular dementia and amyotrophic lateral sclerosis, neurodegeneration abnormal physical conditions; AIDS; such as cardiovascular restenosis, traumatic brain injury, spinal cord injury, cerebral ischemia, cerebral ischemia / perfusion (I / R) injury, Ischemia such as acute and chronic CNS injury ischemia, stroke or myocardial infarction; degenerative diseases of the musculoskeletal system such as osteoporosis; autoimmune diseases such as multiple sclerosis (MS) and type 1 diabetes; and Such as retinal degeneration eye disease.

As the antagonist of IAP, the affinity of the compound of the present invention can be determined using the biological and biophysical test methods described in the examples herein, and the degree of affinity exhibited by the compound can be defined by the term IC ₅₀ value. Specific compounds of the present invention are compounds having an IC ₅₀ value of less than 1 μM, and more particularly less than 0.1 μM.

In one embodiment, the present invention provides a compound for use in the treatment of a disease or abnormal physical condition mediated by IAP (eg, XIAP and / or cIAP (eg cIAP1)). In yet another embodiment, the present invention provides the use of a compound for treating a disease or abnormal physical condition caused by overexpression of IAP (eg, XIAP and / or cIAP (eg, cIAP1)).

In a specific embodiment, the present invention provides the use of a compound for treating an IAP-mediated disease or abnormal physical condition, wherein the compound is used as an antagonist of IAP and is tested in at least one test (e.g., in combination with a displacement test) binding)) The IC ₅₀ of IAP is less than 50 μM. In particular, the IAP is XIAP, cIAP1, and / or cIAP2. In another specific embodiment, the IAP-mediated disease or abnormal physical condition is cancer, which is characterized by overexpressing at least one of IAP and / or 11q22 amplification.

In one specific embodiment, the present invention provides a compound for treating a disease mediated IAP or physical conditions of abnormal, wherein the compound in the test of the IAP (e.g., binding displacement assay) at least one line is less than the IC ₅₀ of IAP 10 μM.

Another embodiment of the present invention provides a compound for use in the manufacture of a medicament for the treatment of an IAP-mediated disease or abnormal physical condition, wherein the compound is an antagonist of IAP and is used in a test (such as in combination with a replacement test ) The IC ₅₀ for at least one IAP is less than 50 μM.

diagnosis method

Before administering a compound of formula (I), the patient must be screened to determine whether the patient has or is likely to have a disease or abnormal physical condition that can be treated with a compound that has an affinity for IAP. The term "patient" includes humans and veterinarians.

For example, a biological sample obtained from a patient can be analyzed to determine whether the disease or abnormal physical condition (eg, cancer) that the patient has or is likely to suffer from is a condition characterized by a genetic or protein abnormality, in which the genetic abnormality Or the abnormal expression of protein leads to the regulation of IAP up-regulation or sensitivity to normal IAP functional pathway or downstream up-regulation of the biochemical pathway of IAP activation.

Examples of abnormalities that cause IAP activation or sensitivity are loss or inhibition of apoptotic pathways, upregulation of receptors or ligands, genetic abnormalities or mutations in receptor or ligand cells. IAP up-regulation of tumors, especially IAP overexpression, can be particularly sensitive to IAP antagonists. For example, a variety of cancers have been found to overexpress XIAP and cIAP, as described in the prior art.

Chromosome 11q22 can be amplified in cell lines, in primary tumors of myeloid cell carcinoma in the esophagus (Imoto et al., 2001) and in the cervix (cervix) (Imoto et al., 2002), as well as in primary tumors. Lung cancer / cell lines (Dai et al., 2003). Immunohistochemistry and western blotting tests have identified both cIAP1 and cIAP2 as potential oncogenes in this region because they are overexpressed in cancers that are not easily amplified.

The term up-regulation includes increased or over-represented performance, including gene amplification (i.e., multiple gene replication numbers), cellular genetic abnormalities, and increased expression due to transcription. Therefore, a diagnostic test can be performed on the patient to detect the marker characteristics of IAP up-regulation. The term diagnosis includes screening. In terms of markers, they include genetic markers, which include measuring DNA composition to detect IAP mutations or 11q22 amplification. The term marker also includes markers of IAP up-regulation features, including the protein quality, protein state, and mRNA amount of the aforementioned proteins.

Diagnostic tests or screening are generally performed on biological samples (i.e., body tissues or fluids), and biological samples are selected from tumor section samples, blood samples (isolated and large amounts of exfoliated tumor cells), cerebrospinal fluid, plasma, serum, saliva , Stool test samples, sputum, chromosome analysis, pleural fluid, ascites, buccal spears, skin sections, or urine.

Methods for detecting and analyzing cytogenetic abnormalities, gene amplification, mutations, and protein up-regulation are known to those skilled in the art. Screening methods may include, but are not limited to, standard methods such as reverse transcriptase polymerase chain reaction (RT-PCR), or in situ hybridization methods such as fluorescent in situ hybridization (FISH).

In RT-PCR screening, the amount of mRNA in a tumor is evaluated by copying the cDNA that produces the mRNA and then amplifying the cDNA by PCR. Methods for PCR amplification, selection of primers, and reaction conditions for amplification are known to those skilled in the art. Nucleic acid manipulation and PCR are performed using general methods, such as Ausubel, FM et al., Eds. (2004) Current Protocols in Molecular Biology, John Wiley & Sons Inc., or Innis, MA et al., Eds. (1990) PCR Protocols: a guide to methods and applications, Academic Press, San Diego. Reactions and manipulations related to nucleic acid technology are also described in Sambrook et al., (2001), 3rd Ed, Molecular Cloning: A Laboratory Manual, Cold Spring Harbor Laboratory Press. Alternatively, commercially available RT-PCR kits (e.g., Roche Molecular Biochemicals) can also be used, or methods such as those described in U.S. Pat. Included in this article for reference.

An example of an in situ hybridization technique used to evaluate mRNA performance may be fluorescent in situ hybridization (FISH) (see Angerer (1987) Meth. Enzymol., 152: 649).

Generally speaking, in situ hybridization includes the following main steps: (1) fixing the tissue to be analyzed; (2) pre-hybridizing the sample to increase the probability of hybridization of the target nucleic acid and reduce non-specific binding; (3) the nucleic acid mixture and Nucleic acids in biological structures or tissues are hybridized; (4) washing after hybridization to remove unlinked hybridized nucleic acid fragments; and (5) detection of hybridized nucleic acid fragments. Probes used in this application are typically labeled with, for example, a radioisotope or a fluorescent reporter. The preferred probe is of sufficient length, for example, about 50, 100, or 200 nucleotides to about 1000 or more nucleotides to specifically hybridize to the target nucleic acid under severe conditions. General methods for performing FISH are described in Ausubel, FM et al., Eds. (2004) Current Protocols in Molecular Biology, John Wiley & Sons Inc and Fluorescence In Situ Hybridization: Technical Overview by John MS Bartlett in Molecular Diagnosis of Cancer, Methods and Protocols, 2nd ed .; ISBN: 1-59259-760-2; March 2004, pps. 077-088; Series: Methods in Molecular Medicine.

The method of gene expression profiling is described in DePrimo et al. (2003), BMC Cancer, 3: 3. Briefly, this method is as follows: a double-stranded cDNA is synthesized from total RNA using the (dT) 24 oligomer used to elicit the first strand of cDMA synthesis, and a second strand of cDNA is synthesized using random hexamer primers. Double-stranded cDNA is used as a template for in vitro cRNA transcription using biotinylated ribonucleotides. The cRNA was chemically cut into fragments using the protocol described in Affymetrix (Santa Clara, CA, USA), and then subjected to overnight hybridization on the Human Genome Array.

Alternatively, the protein products expressed from mRNA can be detected by immunohistochemistry, solid-phase immunoassay using microtiter plates, western blotting methods, two-dimensional SDS-polyacrylamide gel electrophoresis, ELISA, flow cytometry Assays and other methods of detecting specific proteins known in the art are performed. Detection methods may include the use of region-specific antibodies. Those skilled in the art should know that conventional techniques such as detection of IAP up-regulation, detection of IAP mutations or mutations, or detection of 11q22 amplification can be applied in this case.

Abnormal amounts of protein such as IAP can be measured using general protein detection methods, for example, the methods described herein. Increased amounts or overexpression can be detected in tissue samples, for example, the amount of protein in tumor tissue can be measured using, for example, assays from Chemicon International. The target protein can be immunoprecipitated from the sample lysate and its amount can be measured.

Another method for measuring the overexpression or increase in the amount of IAP isomers includes microvessel density measurement. For example, measurement can be performed using the method described by Orre and Rogers (Int J Cancer (1999), 84 (2), 101-8). Detection methods also include the use of markers.

Therefore, these methods can be used to identify tumors that are particularly suitable for treatment with the compounds of the present invention.

Therefore, another aspect of the present invention includes the use of a compound according to the present invention for the manufacture of a medicament for treating or preventing a disease state or an abnormal physical condition of a patient, wherein the patient is considered to have suffered from the disease or screened and screened. A patient with an abnormal physical condition or at risk, and the disease or abnormal physical condition is a disease or condition that can be treated with a compound having an IAP affinity (ie, an IAP antagonist).

Yet another embodiment provides a method of treating a patient suffering from or at risk of a disease or abnormal physical condition (eg, cancer) described herein, comprising administering an effective amount of a compound of formula (I).

Another aspect of the invention includes the use of a compound of the invention for the prevention or treatment of cancer in a patient selected from the group that overexpresses one or more members of the IAP family (e.g., cIAP and / or XIAP) ) Sub-ethnic group.

Another aspect of the present invention includes the use of a compound of the present invention for the prevention or treatment of cancer in a patient selected from a patient suffering from a genetic abnormality in a cell gene that causes IAP to overexpress, for example, From patients suffering from 11q22 amplification.

Vascular normalized MRI measurements (for example, using MRI gradient echo, spin echo, and contrast enhancement methods to measure blood volume, relative vessel size, and vascular permeability) combined with circulating biomarkers can be used to detect the present invention Compound treatment.

Therefore, another aspect of the present invention is a method for diagnosing and treating an IAP-mediated disease state or abnormal physical condition. This method includes (i) screening a patient to determine whether the patient is already or may be suffering. Patients with a disease or abnormal physical condition, wherein the disease or abnormal physical condition can be treated with a compound having an affinity for IAP; and (ii) if the result shows that the patient has the disease or abnormal condition that can be treated, then the patient Compounds of formula (I) and subgroups or examples thereof are administered as described herein.

Drug formula

If the active compound can be administered alone, it is preferably presented as a pharmaceutical composition (eg, a formulation). In a specific embodiment, this is a sterile pharmaceutical composition.

Therefore, the present invention further provides a pharmaceutical composition as described above, and a method for making a pharmaceutical composition, including (eg, mixing) at least one compound of formula (I) (and a second group defined herein) and one or more A pharmaceutically acceptable excipient, and optionally includes other therapeutic or prophylactic agents, as described herein.

Pharmaceutically acceptable excipients may be selected from, for example, carriers (e.g., solid, liquid or semi-solid carriers), adjuvants, diluents, fillers or bulking agents, granulating agents, coatings Agents, controlled release agents, binding agents, disintegrating agents, lubricants, preservatives, antioxidants, buffering agents, suspending agents, thickeners, flavouring agents, sweeteners , Taste masking agent, stabilizer, or any other excipient commonly used in pharmaceutical compositions. Examples of excipients used in various pharmaceutical compositions will be described in detail below.

As used herein, the term "pharmaceutically acceptable" is related to compounds, materials, compositions and / or dosage forms, all of which are within the scope of well-thought-out medical decisions and are applicable to subjects such as, Human), without causing excessive toxicity, irritation, allergic reactions, or other problems or complications, and with reasonable benefit / risk ratios. Each carrier, excipient, etc. must also be "acceptable" and compatible with other ingredients in the formulation.

Pharmaceutical compositions containing compounds of formula (I) can be formulated using known techniques, see, for example, Remington's Pharmaceutical Sciences, Mack Publishing Company, Easton, PA, USA.

The pharmaceutical composition can be any suitable for oral, parenteral, topical, nasal inhalation, bronchial, sublingual, eye, ear, rectal, intravaginal or skin administration. When the composition is used for parenteral administration, it can be formulated for intravenous, intramuscular, intraperitoneal, subcutaneous administration, or delivered directly to the target organ or tissue by injection, perfusion or other administration methods. Delivery can also be by bolus injection, short-term or long-term infusion, and can be via passive delivery or using an appropriate perfusion pump or syringe driver.

Pharmaceutical formulations suitable for parenteral administration include aqueous or non-aqueous sterile injection solutions, which may contain antioxidants, buffer solutions, antibacterial agents, co-solvents, surfactants, organic solvent mixtures, cyclodextrin complexes, emulsifiers ( Used to form and stabilize emulsion formulations), microliposome ingredients to form microlipids, colloidable polymers to form polymeric colloids, cryoprotectants, and, in particular, stabilize the active ingredients in soluble form And a combination of the formulation with an osmotic agent such as blood at the place where it is intended to be received. Pharmaceutical formulations for parenteral administration can also be in the form of aqueous or non-aqueous sterile suspensions, which can include suspensions and thickeners (RG Strickly, Solubilizing Excipients in oral and injectable formulations, Pharmaceutical Research, Vol 21 (2) 2004 , p 201-230).

Formulations can be stored in single or multiple dose containers, such as sealed ampoules, vials, and prefilled injections, and can be stored in a freeze-dried (lyophilised) state, It is only necessary to add a sterile liquid carrier (such as water for injection) immediately before use. In a specific embodiment, the formula is an active pharmaceutical ingredient placed in a bottle, which is reduced in subsequent steps using a suitable diluent.

Pharmaceutical formulations can be prepared by lyophilizing a compound of formula (I) or a subgroup thereof. The lyophilization method refers to a step of freeze-drying a composition. Therefore, freeze-dried and lyophilisation as used herein are synonymous.

Extemporaneous injection solutions and suspensions can be prepared from sterile powders, granules or tablets.

The pharmaceutical composition for parenteral injection of the present invention may also include pharmaceutically acceptable sterile aqueous or non-aqueous solutions, dispersants, suspensions or emulsifiers, and those which are reduced to sterile injection solutions or suspensions just before use. Sterile powder.

Examples of suitable water-soluble or non-water-soluble carriers, diluents, solvents or carriers include water, ethanol, polyols (e.g., glycerol, propylene glycol, polyethylene glycol, and the like), carboxymethyl cellulose, and Suitable mixtures thereof are vegetable oils (such as sunflower oil, safflower oil, corn oil or olive oil), and injectable organic esters (such as ethyl oleate). If proper fluidity must be maintained, for example, by using a thickening or coating substance such as lecithin, maintaining the desired particle size in the case of dispersions, and using a surfactant.

The composition of the present invention may also contain adjuvants such as preservatives, moisturizers, emulsifiers and dispersants. It is also possible to prevent the growth of microorganisms by adding various antibacterial or antifungal agents, such as paraben, chlorobutanol, phenol, sorbic acid and the like. It is also possible to add tonicity-adjusting agents such as sugars, sodium chloride and the like. Long-term absorption of injectable pharmaceutical dosage forms can be achieved by adding agents that delay absorption, such as aluminum monostearate and gelatin.

In a particular embodiment of the present invention, the pharmaceutical composition is a dosage form suitable for intravenous (i.v.) administration (e.g., injection or infusion). For intravenous administration, the solution can be administered by itself or injected into an infusion bag (containing a pharmaceutically acceptable excipient, such as 0.9% physiological saline or 5% glucose) before administration.

In another specific embodiment, the pharmaceutical composition is a dosage form suitable for subcutaneous injection (s.c.) administration.

Pharmaceutical dosage forms suitable for oral administration include tablets (coated or uncoated), capsules (hard or soft shell), round tablets, pills, rhomboid tablets, syrups, solutions, powders, granules, and chitosan ( elixir) and suspensions, sublingual tablets, flakes or patches (eg cheek patches).

Accordingly, the tablet composition may contain a single dose of the active compound and an inert diluent or carrier such as sugars or sugar alcohols such as: lactose, sucrose, sorbitol or mannitol; and / or non-sugar derived diluents such as sodium carbonate , Calcium phosphate, calcium carbonate, or cellulose or derivatives thereof, such as microcrystalline cellulose (MCC), methyl cellulose, ethyl cellulose, hydroxypropyl methyl cellulose, and starches such as corn starch. Tablets can also contain general ingredients, such as polyvinylpyrrolidone as an adhesive and granulator, disintegrants (e.g., swellable crosslinked polymers such as croscarmellose), and lubricants (e.g., Stearate), preservatives (e.g., paraben), antioxidants (e.g., BHT), buffering agents (e.g., phosphate or citrate buffer solutions), and foaming agents (e.g., (Eg, citrate / bicarbonate mixture). Such excipients are known and will not be described in detail here.

Lozenges can be designed to release the drug when in contact with gastric fluid (immediate release of the lozenge), or to control the release of the drug after a long period of time or in contact with a specific area of the gastrointestinal tract (controlled release lozenge).

Capsule formulations can be hard gelatin or soft gelatin types and can contain solid, semi-solid or liquid active ingredients. Gelatin capsules can be made from animal gelatin or its synthetic or plant-derived equivalent.

Solid pharmaceutical forms (eg, tablets, capsules, etc.) can be coated or uncoated. The cover film can be used as a protective film (eg, polymer, wax or varnish), or as a mechanism for controlled release of drugs, or for appearance or identification purposes. A cover film (eg, Eudragit ^™ type polymer) can be designed to release the active ingredient at a predetermined place in the gastrointestinal tract. Therefore, the selected coating must be degraded at a specific pH in the gastrointestinal tract to selectively release the compound in the stomach or ileum, duodenum, jejunum or large intestine.

In addition, if a coating film is not used, or in addition to a coating film, the drug can be presented in the form of a solid matrix, which includes a release control agent, such as a release delaying agent that can be used to control the release of the compound from the gastrointestinal tract. Alternatively, the drug can be provided in a polymer coating film, such as a polymethacrylate polymer coating film, which can be used to selectively release compounds under different acidic or alkaline environments in the gastrointestinal tract. Alternatively, the matrix material or release delay coating can also be in the form of an erodible polymer (e.g., a maleic anhydride polymer) that can be substantially continuously eroded as the agent passes through the gastrointestinal tract. In another aspect, the cover film may be designed to be fragmented through the action of microorganisms in the intestine. In yet another aspect, the active compound can be prepared as a delivery system that can control the release of the compound in an osmotic manner. Osmotic release or other delayed or sustained release formulations (eg, formulations of ion exchange resins) can be prepared according to methods known to those skilled in the art.

The compound of formula (I) can be formulated with a carrier and administered in the form of nano particles. The larger surface area of the nano particles can help their absorption. In addition, nano particles can pass directly through the cell. Nanoparticle drug delivery systems are described in "Nanoparticle Technology for Drug Delivery", edited by Ram B Gupta and Uday B. Kompella, Informa Healthcare, ISBN 9781574448573, published 13 ^th March 2006. Nanoparticles for drug delivery are also described in J. Control. Release, 2003, 91 (1-2), 167-172, and described in Sinha et al., Mol. Cancer Ther. August 1, (2006) 5 , 1909.

The pharmaceutical composition basically includes about 1% (w / w) to about 95% (w / w) active ingredients, and 99% (w / w) to 5% (w / w) of pharmaceutically acceptable excipients Of excipients or excipients. In particular, the composition includes about 20% (w / w) to about 90% (w / w) active ingredients, and 80% (w / w) to 10% of a pharmaceutically acceptable excipient or excipient Of combination. The pharmaceutical composition comprises about 1% to about 95%, especially about 20% to about 90% of the active ingredient. For example, the pharmaceutical composition according to the present invention may be in the form of a single dosage form, such as an ampoule, a vial, a suppository, a pre-filled injection, a dragee, a lozenge, or a capsule.

The choice of pharmaceutically acceptable excipients can be in accordance with the desired physical form of the formulation, for example, it can be selected from diluents (e.g., solid diluents such as fillers and compatibilizers; and liquid diluents such as solvents and co-diluents. Solvents), disintegrating agents, buffering agents, lubricants, flow aids, release control agents (e.g. release retarding or retarding polymers or waxes), adhesives, granulating agents, pigments, plasticizers, antioxidants, preservatives Agent, flavoring agent, taste-masking agent, tonicity adjusting agent and coating agent.

Those skilled in the art can select the proper amount of ingredients to be used in the formulation. For example, tablets and capsules generally contain 0-20% disintegrants, 0-5% lubricants, 0-5% flow aids and / or 0-99% (w / w) fillers or compatibilizers ( (Depending on the drug dose). It may also contain 0 to 10% (w / w) polymer adhesive, 0 to 5% (w / w) antioxidant, and 0 to 5% (w / w) pigment. Sustained-release tablets can additionally contain 0-99% (w / w) release-controlling (eg, delayed) polymers (depending on the dose). Tablets or capsules generally contain 0-10% (w / w) polymers, 0-3% (w / w) pigments, and / or 0-2% (w / w) plasticizers.

Parenteral formulations generally contain 0-20% (w / w) buffer solution, 0-50% (w / w) co-solvent, and / or 0-99% (w / w) water for injection (WFI) ) (Depending on dosage and freeze-drying). Intramuscular depots can also contain 0 to 99% (w / w) oil.

The pharmaceutical composition for oral administration can be obtained by mixing the active ingredient with a solid carrier. If necessary, the resulting mixture is pelletized, and the mixture is processed. If desired or required, appropriate excipients are added. The mixture is then processed into lozenges, dragee cores or capsules. It can also be blended with a polymer or wax matrix to allow the active ingredient to diffuse or release in calculated amounts.

The compounds of the present invention can also be formulated into solid dispersion dosage forms. A solid dispersion dosage form is a homogeneous, finely divided phase of two or more solids. Solid solution (molecular dispersion system), which is one of the forms of solid dispersion, is commonly used in pharmaceutical technology (see Chiou and Riegelman, J. Pharm. Sci., 60, 1281-1300 (1971)) and can be used in Increase the solubility of poorly water-soluble drugs and increase bioavailability.

The present invention also provides a solid pharmaceutical form including the aforementioned solid solution. Solid pharmaceutical forms include lozenges, capsules, chewable lozenges, and dispersible or foaming lozenges. Known excipients can be mixed with a solid solution to provide the desired pharmaceutical form. For example, a capsule may contain a solid solution blended with (a) a disintegrant and a lubricant, or (b) a disintegrant, a lubricant, and a surfactant. In addition, the capsule may contain a compatibilizer such as lactose or microcrystalline cellulose. Lozenges may contain a solid solution blended with at least one of a disintegrant, a lubricant, a surfactant, a compatibilizer, and a glidant. The chewable tablet may contain a solid solution mixed with a compatibilizer and a lubricant, and if desired, an additional sweetener (such as an artificial sweetener), and an appropriate flavoring agent may be added. A solid solution can also be formed by spraying a drug solution with an appropriate polymer on the surface of an inert carrier such as sugar beads (non-pareils). These sugar beads can then be filled in capsules or compressed into tablets.

The pharmaceutical formula can be presented to the patient in the form of a "patient pack", the complete package of which is included in a single package, and is usually a blister pack. Patient kits have advantages over traditional prescriptions. In traditional prescriptions, pharmacists divide patient supply from the total supply, but the advantage of patient kits is that patients must see the package insert in the kit. This point is generally missing from patient prescriptions. The kit contains instructions for the drug to make it easier for the patient to follow the instructions of the doctor.

Compositions for topical and nasal delivery include ointments, creams, sprays, patches, gels, droplets, and implants (e.g., intraocular inserts). This composition can be formulated in a conventional manner.

Examples of rectal or intravaginal formulations include pessaries and suppositories, for example, which may be formed from a moldable or waxy material containing an active compound. The active compound solution can also be used for rectal administration.

The composition to be administered by inhalation may be an inhaled powder composition, a liquid, or a powder spray, and may be administered in a general form such as a powder inhalation device or an aerosol dispersion device. These devices are known. For administration by inhalation, powder formulations generally include active drugs and inert solid powder diluents such as lactose.

Compounds of formula (I) will generally be presented in a single dosage form, and as such will generally contain a sufficient amount of the compound to provide the desired biological activity. For example, the formulation may contain 1 ng (nanogram) to 2 g (gram) of active ingredient, such as 1 ng to 2 mg (milligram) of active ingredient. Within this range, special sub-ranges of the compound are 0.1 mg to 2 g of active ingredient (typically more commonly 10 mg to 1 g, such as 50 mg to 500 mg), or 1 μg (microgram) to 20 mg (e.g. , 1 μg to 10 mg, such as 0.1 mg to 2 mg of active ingredient).

In oral compositions, a single dosage form may contain from 1 mg to 2 g, typically more commonly from 10 mg to 1 g, such as from 50 mg to 1 g, such as from 100 mg to 1 g of active compound.

The active compound can be administered to a desired patient (e.g., a human or animal patient) in a dose sufficient to achieve the desired therapeutic effect.

treatment method

The compounds of formula (I) and their subgroups as defined herein can be used to prevent or treat many disease states or abnormal physical conditions mediated by IAP. Therefore, according to another aspect of the present invention, a method for treating a disease state or abnormal physical condition (such as cancer) mediated by IAP (eg, XIAP and / or cIAP) is provided, including the subject to be administered. A compound of formula (I) as described herein is administered. According to yet another aspect of the present invention, a method is provided for treating a disease state or abnormal physical condition (such as cancer) overexpressed by IAP (eg, XIAP and / or cIAP), including administering a subject to be administered as described herein. The compound of formula (I). Examples of such disease states and abnormal physical conditions are as previously described, and specifically include cancer.

The compound is generally administered to a subject to be administered, such as a human or animal patient, especially a human.

Compounds are generally administered in amounts that are therapeutically or prophylactically effective and generally do not cause toxicity. However, in some cases (e.g., life-threatening diseases), the benefits of administering a compound of formula (I) are more important than the disadvantages of any toxic reaction and side effects. In this case, it can be considered that administration will cause a certain degree of Amount of toxic compounds.

The compounds can be administered over a long period of time to maintain an effective therapeutic effect, or only for a short period of time. Alternatively, it may be administered continuously or intermittently (eg, periodically).

Although higher or lower doses can be administered as required, the general daily dose of the compound of formula (I) can be 100 pg (picogram) to 100 mg per kilogram of body weight, and usually 5 ng to 25 mg per kilogram of body weight, And it is usually 10 ng to 15 mg per kilogram of body weight (eg, 10 ng to 10 mg per kilogram, and usually 1 μg to 20 mg per kilogram, for example, 1 μg to 10 mg per kilogram). For example, the compound of formula (I) can be administered daily or every two, or three, or four, or five, or six, or seven, or ten, or fourteen, or twenty-one, or twenty-eight or twenty-eight repeating cycles.

The oral dose range of the compound of the present invention is, for example, 1 to 1500 mg, 2 to 800 mg, or 5 to 500 mg, such as 2 to 200 mg or 10 to 1000 mg, and specific examples of the dose include 10, 20, 50, and 80 mg. The compounds can be administered once a day or more than once. The compound can be administered in a continuous manner (ie, taken continuously throughout the course of the treatment). Alternatively, the compound may be administered intermittently (i.e., continuously for a period of time (e.g., one week) during the entire treatment course, and then discontinued for a period of time (e.g., one week), and then continued for another period of time (e.g., one week) Wait). Examples of treatment courses using intermittent dosing include: one week dosing and one week dosing; two weeks dosing and one week dosing; two weeks dosing and one week dosing; and two weeks dosing and two week dosing; Or a two-week suspension period with four weeks of dosing; or a three-week suspension period with one week of dosing and one or more cycles, such as 2, 3, 4, 5, 6, 7, 8, 9, or 10 or more Cycles.

In a specific dose schedule, the compound of formula (I) is administered to a patient for one hour per day for up to ten days, particularly up to five days per week, and the treatment is repeated at predetermined intervals, such as two to four weeks, especially for Every three weeks.

More particularly, the compound of formula (I) is administered to a patient for one hour per day for five days, and the treatment is repeated every three weeks.

In another specific dose schedule, the patient is perfused for 30 minutes to 1 hour, and then maintenance infusion is administered for a period of time, such as 1 to 5 hours (eg, 3 hours).

In yet another specific dose schedule, the patient is administered continuous perfusion for 12 hours to 5 days, especially for 24 to 72 hours.

In another specific dosage schedule, the compound is administered to a patient orally once a week.

In another specific dosage schedule, the compound is administered to a patient orally once a day for between 7 and 28 days, such as 7, 14, or 28 days.

In another specific dosage schedule, the compound is administered orally to a patient once a day for 1 day, 2 days, 3 days, 5 days, or 1 week, and then the drug is discontinued for a predetermined number of days to complete a one or two week cycle .

In another specific dosage schedule, the compound is administered orally once a day to the patient for 2 weeks, and then the drug is discontinued for 2 weeks.

In another specific dose schedule, the compound is administered orally once a day to the patient for 2 weeks, and then the drug is discontinued for 1 week.

In another specific dose schedule, the compound is administered orally once a day to a patient for one week, and then the drug is discontinued for one week.

However, the amount of the compound to be administered and the form of the composition to be used will ultimately be related to the physical characteristics of the disease or physiological abnormality to be treated, and the judgment of the physician.

It has been found that IAP antagonists can be used as a single agent or in combination with other anticancer agents. For example, it is beneficial to combine an antagonist that induces apoptosis with another agent that regulates cell growth through different mechanisms, so as to treat the two symptoms of cancer development. Consolidation experiments are described in Chou TC, Talalay P. Quantitative analysis of dose-effect relationships: the combined effects of multiple drugs or enzyme inhibitors. Adv Enzyme Regulat 1984; 22: 27-55.

The compounds defined in the present invention can be administered as a single therapeutic agent, or can be administered in combination with one or more other compounds (or therapies) to treat a specific disease state, for example, the cancerous tumor diseases described above. In the treatment of the aforementioned abnormal physical conditions, the compounds of the present invention can be advantageously used in combination with one or more other pharmaceutical agents, especially in the treatment of cancer with other anticancer agents or adjuvants (therapy adjuvants). Examples of other therapeutic agents or therapies that can be administered with a compound of formula (I) (whether at the same time or at different time intervals) include, but are not limited to: a topoisomerase I inhibitor; an anti-metabolic drug; a tubulin-targeted agent DNA binding and topoisomerase II inhibitors; methylation reagents; monoclonal antibodies; antihormonal reagents; signal transmission inhibitors; protein body inhibitors; DNA methyltransferase inhibitors; cytokines and retinoids Retinoids; chromatin-targeted therapies; radiotherapy; and other therapeutic or preventive agents.

Specific examples of anticancer agents or adjuvants (or their salts) include, but are not limited to, any agent selected from the group (i)-(xlvi) and the selective group (xlvii):

(i) a platinum compound, such as cisplatin (optionally used in combination with amifostine), carboplatin, or oxaliplatin;

(ii) taxane compounds, such as paclitaxel, paclitaxel protein-binding particles (Abraxane ^™ ), docetaxel, cabazitaxel, or larotaxel;

(iii) a topoisomerase I inhibitor, such as a camptothecin compound, such as camptothecin, irinotecan (CPT11), SN-38, or topotecan;

(iv) a topoisomerase II inhibitor, such as an antitumor epipodophyllotoxin or podophyllotoxin derivative, such as etoposide, or teniposide;

(v) Vinca alkaloids, such as vinblastine, vincristine, liposomal vincristine (Onco-TCS), vinorelbine, vinorelbine Vindesine, vinflunine or vinvesir;

(vi) nucleoside derivatives, such as 5-fluorouracil (5-FU, optionally combined with leucovorin), gemcitabine, capecitabine, tega Tegafur, UFT, S1, cladribine, cytarabine (Ara-C, cytosine arabinoside), fludarabine, clofarabine, or naira Shore (nelarabine);

(vii) Anti-metabolic drugs, such as clofarabine, aminopterin, methotrexate, azacitidine, cytarabine, fluorouridine (floxuridine), pentostatin, thioguanine, thiopurine, 6-mercaptopurine, or hydroxyurea (hydroxycarbamide);

(viii) alkylating agents, such as nitrogen mustards or nitrosourea, such as cyclophosphamide, chlorambucil, carmustine, BCNU), bendamustine, thiotepa, melphalan, treosulfan, lomustine (CCNU), altretamine Busulfan, dacarbazine, estramustine, fotemustine, ifosfamide (optionally combined with mesna) (Used in), piperobromine (pipobroman), procarbazine, streptozocin, temozolomide, uracil, mechlorethamine, methylcyclohexylchloroethyl Methylcyclohexylchloroethylnitrosurea, or nimustine (ACNU);

(ix) anthracyclines, anthracenediones, and related drugs, such as daunorubicin, doxorubicin (which is selectively combined with dexrazoxane Used), doxorubicin microliposome formulations (e.g., Caelyx ^TM , Myocet ^TM , Doxil ^TM ), idarubicin, mitoxantrone, epirubicin, amine benzene Acridine (amsacrine), or valrubicin;

(x) Epothilones, such as ixabepilone, patupilone, BMS-310705, KOS-862 and ZK-EPO, epothilone A , Epothilone B, desoxyepothilone B (also known as epothilone D or KOS-862), aza epothilone B (also known as BMS-247550), aulimalide, isolaulimalide, or luetherobin;

(xi) a DNA methyltransferase inhibitor, such as temozolomide, azacytidine or decitabine, or SGI-110;

(xii) antifolates, such as methotrexate, pemetrexed disodium, or raltitrexed;

(xiii) cytotoxic antibiotics, such as antinomycin D, bleomycin, mitomycin C, dactinomycin, carminomycin, Daunomycin, levamisole, plicamycin, or mithramycin;

(xiv) a tubulin binding agent, such as combestatin, colchicines, or noeodazole;

(xv) Signal transduction inhibitors, such as kinase inhibitors (e.g., EGFR (endothelial growth factor receptor) inhibitors, VEGFR (vascular endothelial growth factor receptor) inhibitors, PDGFR (platelet-derived growth factor receptor) ) Inhibitors, MTKI (multi-standard kinase inhibitors), Raf inhibitors, mTOR inhibitors, such as imatinib mesylate, erlotinib, gefitinib, Dasatinib, lapatinib, dovotinib, axitinib, nilotinib, vandetanib, vata Vatalinib, pazopanib, sorafenib, sunitinib, temsirolimus, everolimus (RAD 001), Vemurafenib (PLX4032 / RG7204), dabrafenib, encorafenib, or IκB kinase inhibitors, such as SAR-113945, bardoxolone, BMS-066, BMS-345541, IMD-0354, IMD -2560, or IMD-1041, or MEK inhibitors, such as Selumetinib (AZD6244) and Trametinib (GSK121120212);

(xvi) aurora kinase inhibitors, such as AT9283, barasertib (AZD1152), TAK-901, MK0457 (VX680), cenisertib (R-763), danusertib ) (PHA-739358), alisertib (MLN-8237), or MP-470;

(xvii) CDK inhibitors, such as AT7519, roscovitine, seliciclib, alvocidib (flavopiridol), dinasicilib (dinaciclib) (SCH-727965), 7-hydroxy-staurosporine (UCN-01), JNJ-7706621, BMS-387032 (also known as SNS-032), PHA533533, PD332991 , ZK-304709, or AZD-5438;

(xviii) PKA / B inhibitors and PKB (akt) pathway inhibitors, such as KRX-0401 (perifosine / NSC 639966), ipatasertib (GDC-0068; RG-7440), afuresertib (GSK- 2110183; 2110183), MK-2206, MK-8156, AT13148, AZD-5363, triciribine phosphate (VQD-002; triciribine phosphate monohydrate) (API-2; TCN-P; TCN-PM; VD-0002), RX-0201, NL-71-101, SR-13668, PX-316, AT13148, AZ-5363, Semaphore, SF1126, or Enzastaurin HCl (LY317615), or MTOR Inhibitors, such as rapamycin analogs, such as RAD 001 (everolimus), CCI 779 (temsirolemus), AP23573, and ridaforolimus, sirolimus ( sirolimus) (originally known as rapamycin), AP23841 and AP23573, calmodulin inhibitors such as CBP-501 (forkhead translocation inhibitors), enzastaurin HCl ( LY317615), or PI3K inhibitors such as dactolisib (BEZ235), buparisib (BKM-120; NVP-BKM-120), BYL719 Copanlisib, (BAY-80-6946), ZSTK-474, CUDC-907, apitolisib (GDC-0980; RG-7422), pictilisib (pictrelisib, GDC-0941, RG-7321 ), GDC-0032, GDC-0068, GSK-2636771, idelalisib (previously called CAL-101, GS 1101, GS-1101), MLN1117 (INK1117), MLN0128 (INK128), IPI-145 ( INK1197), LY-3023414, ipatasertib, afuresertib, MK-2206, MK-8156, LY-3023414, LY294002, SF1126 or PI-103, or sonolisib (PX-866);

(xix) Hsp90 inhibitors such as AT13387, herbimycin, geldanamycin (GA), 17-acrylamido-17-demethoxygeldanamycin (17 -allylamino-17-desmethoxygeldanamycin) (17-AAG), such as NSC-330507, Kos-953 and CNF-1010, 17-dimethylaminoethylamino-17-desmethoxygeldanamycin hydrogen 17-dimethylaminoethylamino-17-demethoxygeldanamycin hydrochlorid (17-DMAG), such as NSC-707545 and Kos-1022, NVP-AUY922 (VER-52296), NVP-BEP800, CNF-2024 (BIIB-021, oral purine) ), Ganetespib (STA-9090), SNX-5422 (SC-102112), or IPI-504;

(xx) Monoclonal antibodies (unconjugated or conjugated to radioisotopes, toxins or other agents), antibody derivatives and related agents, such as anti-CD, anti-VEGFR, anti-HER2, anti- CTLA4, anti-PD-1, or anti-EGFR antibodies, such as rituximab (CD20), ofatumumab (CD20), ibritutumab (tiuxetan) (CD20) , GA101 (CD20), tositumomab (CD20), epratuzumab (CD22), lintuzumab (CD33), gemtuzumab ozogamicin (CD33), alemtuzumab (CD52), galiximab (CD80), trastuzumah (HER2 antibody), pertuzumab (pertuzumab) ( HER2), trastuzumab-DM1 (HER2), ertumaxomab (HER2 and CD3), cetuximab (EGFR), panitumumab (EGFR) , Necitumumab (EGFR), nimotuzumab (EGFR), bevacizumab (VEGF), catumaxumab (EpCAM and CD3), Abagovomab (CA125 ), Farletuzumab (folate receptor), elotuzumab (CS1), denosumab (RANK ligand), figitumumab (IGF1R), CP751,871 (IGF1R), mapatumumab (TRAIL receptor), metMAB (met), mitumomab (GD3 ganglioside), naptumomab estafenatox ) (5T4), siltuximab (IL6), or an immunomodulator such as a CTLA-4 blocking antibody and / or an antibody against PD-1 and PD-L1 and / or PD-L2, For example, ipilimumab (CTLA4), MK-3475 (pembrolizumab, formerly known as lambrolizumab, anti-PD-1), nivolumab (anti-PD- 1), BMS-936559 (anti-PD-L1), MPDL320A, AMP-514 or MEDI4736 (anti-PD-L1), or trimelimumab (formerly known as ticilimumab), CP-675,206, anti-CTLA-4);

(xxi) estrogen receptor antagonists or selective estrogen receptor modulators (SERMs) or inhibitors of estrogen synthesis, such as tamoxifen, fulvestrant, toremifene ( toremifene), droloxifene, faslodex, or raloxifene;

(xxii) aromatase inhibitors and related drugs, such as exemestane, anastrozole, letrazole, testolactone aminoglutethimide, rice Mitotane or vorozole;

(xxiii) antiandrogens (ie, androgen receptor antagonists) and related agents, such as, bicalutamide, nilutamide, flutamide, cyclopropane Progesterone (cyproterone) or ketoconazole;

(xxiv) hormones and their analogs, such as medroxyprogesterone, diethylstilbestrol (also known as diethylstilboestrol) or octreotide;

(xxv) steroids, such as drostanolanol propionate, megestrol acetate, nandrolone (decanoate, phenpropionate) , Fluoxymestrone, or gossypol;

(xxvi) a steroid chromosome P450 17α-hydroxylase-17,20-lyase inhibitor (CYP17), such as abiraterone;

(xxvii) Gonadotropin-releasing hormone agonists or antagonists (GnRA), such as abarelix, goserelin acetate, histrelin acetate , Leuprolide acetate, triptorelin, buserelin, or deslorelin;

(xxviii) Glucocorticoids, such as prednisone, prednisolone, and dexamethasone;

(xxix) Differentiating agents, such as retinoid, rexinoid, vitamin D or vitamin A acid (retinoic acid) and vitamin A acid metabolism blocking agents (RAMBA), such as isotretinoin ( accutane), alitretinoin, bexarotene, or tretinoin;

(xxx) Farnesyltransferase inhibitors, such as tipifarnib;

(xxxi) Chromatin-targeted therapies such as histone deacetylase (HDAC) inhibitors, such as panobinostat, resminostat, abexinostat, vorino He (vorinostat), romidepsin, belinostatin, entinostat, quisinostat, pracinostat, tefinostat, mocetinostat, gee Givinostat, CUDC-907, CUDC-101, ACY-1215, MGCD-290, EVP-0334, RG-2833, 4SC-202, romidepsin, AR-42 (Ohio State University ), CG-200745, valproic acid, CKD-581, sodium butyrate, suberoylanilide hydroxamide acid (SAHA), depsipeptide (FR 901228), dacinostat (NVP-LAQ824), R306465 / JNJ-16241199, JNJ-26481585, trichostatin A, chlamydocin, A-173, JNJ -MGCD-0103, PXD-101, or apicidin;

(xxxii) proteosome inhibitors, such as bortezomib, carfilzomib (CEP-18770), ixazomib (MLN-9708), oprozomib (ONX-0912) , Or marizomib;

(xxxiii) a photodynamic therapeutic agent, such as porfimer sodium or temoporfin;

(xxxiv) marine organism-derived anticancer agents, such as trabectidin;

(xxxv) radiation-calibrating drugs used for radioimmunotherapy, for example, labeled with beta particles releasing isotopes (eg, iodine-131, yttrium-90), or alpha particles releasing isotopes (eg, bismuth-213 or thallium-225), For example, ibritumomab, or Iodine tositumomab;

(xxxvi) a telomerase inhibitor, such as telomestatin;

(xxxvii) a matrix metalloproteinase inhibitor, such as batimastat, marimastat, prinostat, or metastat;

(xxxviii) recombinant interferon (eg, interferon-γ and interferon α) and interleukin (eg, interleukin 2), such as aldesleukin, denileukin diftitox, interferon α2a, interferon α2b, or polyethylene glycol interferon α2b (peginterferon alfa 2b);

(xxxix) a selective immune response modifier, such as thalidomide, or lenalidomide;

(xl) a therapeutic vaccine, such as sipuleucel-T (Provenge) or OncoVex;

(xli) cytokine activators, including Picibanil, Romurtide, Sizofiran, Virulizin, or Thymosin;

(xlii) Arsenic trioxide;

(xliii) a G-protein coupled receptor (GPCR) inhibitor, such as atrasentan;

(xliv) enzymes, such as L-asparaginase, pegaspargase, rasburicase, or pegademase;

(xlv) a DNA repair inhibitor, such as a PARP inhibitor, such as olaparib, velaparib, iniparib, INO-1001, AG-014699, or ONO-2231;

(xlvi) death receptor agonists (e.g., TNF-related apoptosis-inducing ligand (TRAIL) receptors), e.g., mapatumumab (formerly HGS-ETR1), kanamu Monoclonal antibody (conatumumab) (previously known as AMG 655), PRO95780, lexatumumab, dulanermin, CS-1008, apomorph, or recombinant TRAIL ligand, such as Recombinant human TRAIL / Apo2 ligand;

(xlvii) preventive agents (adjuvants); that is, agents that reduce or mitigate certain side effects associated with chemotherapy, such as:-anti-emetic agents;-prevent or reduce chemotherapeutic related neutropenia Agents for the continued occurrence of neutropenia and the prevention of complications caused by declining platelets, red blood cells, or white blood cells, such as interleukin-11 (e.g., oprelvekin), erythropoietin, EPO) and its analogs (eg, darbepoetin alfa), colony-stimulating factor analogs, such as granulocyte macrophage-community stimulating factor (GM-CSF) (e.g., sargramostim), and granule-community stimulating factor (G-CSF) and its analogs (e.g., filgrastim, PEGylated filgrastim) Pegfilgrastim);-inhibitors of bone resorption, such as denosumab or bisphosphonates, such as zoledronate, zoledronic acid, Pamidronate and ibandronate );-Agents that inhibit inflammation, such as dexamethasone, prednisone, and prednisolone;-to reduce cancer in patients with acromegaly or other rare hormones Reagents for the growth hormone and IGF-1 (and other hormones) in blood, such as the synthetic form of hormone somatostatin, such as octreotide acetate;-a drug detoxifier that reduces the amount of folic acid, For example, leucovorin, or folinic acid;-pain agents, such as opiates, such as morphine, diamorphine, and fentanyl (fentanyl);-non-steroidal anti-inflammatory drugs (NSAID), such as COX-2 inhibitors, such as celecoxib, etoricoxib, and lumiracoxib;-for mucositis Agents, for example, palifermin;-agents for treating side effects such as anorexia, cachexia, oedema, or thromboembolic episodes, such as megestrol acetate .

In a specific embodiment, the anticancer agent is selected from the group consisting of recombinant interferon (such as interferon-γ and interferon α) and interleukin (for example, interleukin 2), for example, aldileukin, interleukin fusion toxin, Interferon α2a, interferon α2b, or pegylated interferon α2b; interferon α2 (500 μ / ml), especially interferon β; and messaging inhibitors such as kinase inhibitors (for example, EGFR (endothelial growth Factor receptor) inhibitors, VEGFR (vascular endothelial growth factor receptor) inhibitors, PDGFR (platelet-derived growth factor receptor) inhibitors, MTKI (multi-standard kinase inhibitors), Raf inhibitors, mTOR inhibitors, such as, Imatinib mesylate, erlotinib, gefitinib, dasatinib, lapatinib, dovotinib , Axitinib, nilotinib, vandetanib, vatalinib, pazopanib, sorafenib, Shu Sunitinib, temsirolimus, everolimus (RAD 001), vemura fenib) (PLX4032 / RG7204), dabrafenib, encorafenib, or IκB kinase inhibitors, such as SAR-113945, bardoxolone, BMS-066, BMS-345541, IMD-0354, IMD-2560, or IMD -1041, or MEK inhibitors, such as Selumetinib (AZD6244) and Trametinib (GSK121120212), especially Raf inhibitors (e.g., vemurafenib) or MEK Inhibitors (e.g. trametinib).

Each compound in the combination of the present invention can be administered individually in different doses and through different routes. As such, the dosage of each drug in two or more agents may be different: each drug may be administered at the same time or at different times. Those skilled in the art can know the dosage and combination therapy to be used through their general general knowledge. For example, a compound of the invention may be used in combination with one or more other agents and administered according to a combination therapy in which such agents already exist. Examples of general concomitant therapies are provided below.

The taxane compounds are preferably administered at a dose of 50 to 400 mg (mg / m ² ) per square meter of body surface area (e.g., 75 to 250 mg / m ² ) in each course of treatment, especially the taxol dose. About 175 to 250 mg / m ² , and the dosage of docetaxel is about 75 to 150 mg / m ² .

The camptothecin compound is preferably administered at a dose of 0.1 to 400 mg (mg / m ² ) per square meter of body surface area (e.g., 1 to 300 mg / m ² ) in each course of treatment, especially The dose of irinotecan is about 100 to 350 mg / m ² , and the dose of topotecan is about 1 to 2 mg / m ² .

An anti-tumor podophyllotoxin derivative is preferably administered at a dose of 30 to 300 mg (mg / m ² ) per square meter of body surface area (e.g., 50 to 250 mg / m ² ) during each course of treatment. In particular, the dose of etoposide is about 35 to 100 mg / m ² , and the dose of podophyllotoxin (teniposide) is about 50 to 250 mg / m ² .

The anti-tumor vinca alkaloid is preferably administered at a dose of 2 to 30 mg (mg / m ² ) per square meter of body surface area, especially the vinblastine dose. About 3 to 12 mg / m ² , the dose of vincristine is about 1 to 2 mg / m ² , and the dose of vinorelbine is about 10 to 30 mg / m ² .

The antitumor nucleotide derivative is preferably administered at a dose of 200 to 2500 mg (mg / m ² ) (for example, 700 to 1500 mg / m ² ) at a body surface area per square meter in each course of treatment, particularly The 5-FU dose is about 200 to 500 mg / m ² , the gemcitabine dose is about 800 to 1200 mg / m ² , and the capecitabine dose is about 1000 to 2500 mg / m ² .

For example, alkylating agents such as nitrogen mustards or nitrosourea are preferably administered at a dose of 100 to 500 mg (mg / m ² ) per square meter of body surface area during each course of treatment. (Eg 120 to 200 mg / m ² ), especially cyclophosphamide at a dose of about 100 to 500 mg / m ² , chlorambucil at a dose of about 0.1 to 0.2 mg / m ² , carmus The carmustine dose is about 150 to 200 mg / m ² , while the lomustine dose is about 100 to 150 mg / m ² .

Antitumor anthracycline derivatives are preferably administered at a dose of 10 to 75 mg (mg / m ² ) per square meter of body surface area (e.g., 15 to 60 mg / m ² ) during each course of treatment. In particular, the dose of doxorubicin is about 40 to 75 mg / m ² , the dose of daunorubicin is about 25 to 45 mg / m ² , and the dose of idarubicin is about 10 to 15 mg / m ² .

The administration of the anti-estrogen agent depends on the special agent and the abnormal physical condition to be treated, and it is preferably administered at a dose of 1 to 100 mg per day. Tamoxifen is preferably administered orally, its dosage is 5 to 50 mg, especially 10 to 20 mg, taken twice a day, and continued treatment for a sufficient time to achieve or maintain the therapeutic effect. Toremifene is preferably administered orally, its dose is about 60 mg, and it is taken once a day, and the treatment is continued for a sufficient time to achieve or maintain the therapeutic effect. Anastrozole is preferably administered orally, its dose is about 1 mg, and it is taken once a day. Droloxifene is preferably administered orally, its dosage is about 20-100 mg, and it is taken once a day. Raloxifene is preferably administered orally, its dose is about 60 mg, and it is taken once a day. Exemestane is preferably administered orally, at a dose of about 25 mg, and taken once a day.

The antibody is preferably administered at a dose of about 1 to 5 mg (mg / m ² ) per square meter of body surface area, or if it is to be changed, it can be administered in a dosage manner known in the art. Trastuzumab is preferably administered at a dose of 1 to 5 mg (mg / m ² ) per square meter of body surface area during each course of treatment, especially at 2 to 4 mg / m ² . Dosing.

When the compound of formula (I) is combined with one, two, three, four or more (preferably one or two, and more particularly one) other therapeutic agents, the compounds may be administered simultaneously or sequentially. In the case of sequential administration, two or more compounds can be administered in a certain amount and in a manner of administration over a period of time, which is sufficient to ensure that a better or synergistic effect can be achieved. When administered sequentially, they can be administered at relatively short intervals (e.g., about 5 to 10 minutes) or longer intervals (e.g., 1, 2, 3, 4 or more hours apart, or as needed) (Can be separated for a longer period of time), and the precise dose of treatment must be consistent with the characteristics of the therapeutic agent. In each course, the dosage of the medicament may be administered, for example, once, twice, or multiple times, and the administration may be repeated, for example, every 7, 14, 21, or 28 days.

In a specific embodiment, a compound of formula (I) is provided for the manufacture of a medicine for treatment, wherein the compound is used in combination with one, two, three, or four other therapeutic agents. In another embodiment, a medicine for treating cancer is provided, which comprises a compound of formula (I), wherein the medicine is used in combination with one, two, three, or four other therapeutic agents. The present invention further provides the use of a compound of formula (I) for the manufacture of a medicine that enhances the response rate of cancer patients being treated with one, two, three, or four other therapeutic agents or promotes the response rate.

It should be understood that the particular method and sequence of administration, as well as the individual doses and treatments of each component in the combined treatment, are related to the specific other pharmaceutical agents and compounds of the present invention to be administered, their route of administration, the specific tumor to be treated, and Specific to the subject to be treated. Those skilled in the art can use conventional methods and refer to the information provided herein to easily determine the optimal method and order of administration, as well as the dosage and treatment.

When combining, one skilled in the art can determine the weight ratio of the compound according to the invention to one or more other anticancer agents. It is well known to those skilled in the art that the dosage ratio, actual dosage and frequency are according to the specific compounds of the present invention and other anticancer agents used, specific abnormal physical conditions to be treated, severity of the conditions to be treated, age, weight, sex, diet , The time of administration, and the usual physiological condition of a particular patient, and it is related to the way of administration and other drugs that the individual may be taking. Furthermore, it is clear that the effective daily dose can be increased or decreased depending on the response of the subject being treated and / or the assessment of the physician prescribing the compound of the invention. The specific weight ratio of the compound of formula (I) of the present invention to other anticancer agents may be in the range of 1/10 to 10/1, more specifically in the range of 1/5 to 5/1, and more specifically in 1/3. To the range of 3/1.

The compounds of the present invention can also be administered with non-chemotherapy, such as radiotherapy, photodynamic therapy, gene therapy; surgery and diet control.

The compounds of the present invention can also be used in therapeutic applications that sensitize tumor cells to radiotherapy and chemotherapy. Therefore, the compounds of the present invention can be used as "radiosensitizers" and / or "chemical sensitizers" or can be administered in combination with other "radiosensitizers" and / or "chemical sensitizers". In a specific embodiment, the compound of the present invention is used as a chemical sensitizer.

The term "radiosensitizer" refers to the administration of a therapeutically effective amount of a molecule to a patient to increase the sensitivity of the cells to ionizing radiation and / or to enhance the therapeutic effect of diseases that can be treated with ionizing radiation.

The term "chemical sensitizer" refers to the administration of a therapeutically effective amount of a molecule to a patient to increase the sensitivity of the cells to chemotherapy and / or to enhance the effect of a disease that can be treated with chemotherapy.

In a specific embodiment, the compound of the present invention is administered with a "radiosensitizer" and / or a "chemical sensitizer". In a specific embodiment, the compound of the present invention is administered with an "immunosensitizer".

The term "immunosensitizer" refers to the administration of a therapeutically effective amount of a molecule to a patient to increase the sensitivity of a cell to an IAP antagonist, for example, by initiating the release of TNF to enhance or increase the immune response.

Many cancer treatment courses today use radiosensitizers with X-rays. Examples of X-ray activated radiosensitizers include, but are not limited to, the following: metronidazole, misonidazole, desmethylmisonidazole, pimonidazole ), Etanidazole, nimorazole, mitomycin C, RSU 1069, SR 4233, EO9, RB 6145, nicotinamide, 5-bromodeoxy 5-bromodeoxyuridine (BUdR), 5-iododeoxyuridine (IUdR), bromodeoxycytidine, fluorodeoxyuridine (FudR), hydroxyurea, cis Platinum, and therapeutically effective analogs and their derivatives.

Photodynamic therapy of cancer (PDT) is a radiation initiator that uses visible light as a sensitizer. Examples of photodynamic radiosensitizers include the following, but are not limited to: hematoporphyrin derivatives, photofrin, benzoporphyrin derivatives, tin etioporphyrin, Pheoborbide-a, bacteriochlorophyll-a, naphthalocyanines, phthalocyanines, zinc phthalocyanine, and treatment Effective analogs and their derivatives.

Radiosensitizers can be administered with one or more therapeutically effective amounts of other compounds, including other compounds including, but not limited to: compounds of the invention; compounds that enhance the incorporation of radiosensitizers into target cells; control therapeutic agents , Nutrients, and / or compounds that oxygen flows to target cells; chemotherapeutic agents that require or do not require additional radiation to act on tumors; or other therapeutically effective compounds for the treatment of cancer or other diseases.

Chemical sensitizers can be administered with one or more therapeutically effective amounts of other compounds, including other compounds including, but not limited to: compounds of the invention; compounds that enhance the incorporation of chemical sensitizers into target cells; and control of therapeutic agents , Nutritional agents, and / or compounds that oxygen flows to target cells; chemotherapeutic agents that act on tumors or other compounds that are effective in treating cancer or other diseases. Calcium antagonists, such as verapamil, have been shown to be used in combination with anti-tumor agents to produce chemosensitivity in tumor cells that are resistant to the received chemotherapeutic agent, and to make these compounds resistant to Drug-sensitive tumors produce efficacy.

Examples of immunosensitizers include the following, but are not limited to, immunomodulators, such as monoclonal antibodies, such as immune checkpoint antibodies [eg, CTLA-4 blocking antibodies and / or anti-PD-1 and PD-L1 and / Or PD-L2 antibodies, such as ipilimumab (CTLA4), MK-3475 (pembrolizumab, formerly known as lambrolizumab, anti-PD-1), nivolumab (nivolumab) (anti-PD-1), BMS-936559 (anti-PD-L1), MPDL320A, AMP-514 or MEDI4736 (anti-PD-L1), or trimelimumab (previously known as replacement Ticilimumab, CP-675,206, anti-CTLA-4); or messaging inhibitors; or cytokines (eg, recombinant interferon); or oncolyte virus; or immune adjuvants (E.g. BCG).

Immune sensitizers can be administered with one or more therapeutically effective amounts of other compounds, including other compounds including, but not limited to: compounds of the invention; compounds that enhance the incorporation of immune sensitizers into target cells; control therapeutic agents , Nutritional agents, and / or oxygen compounds to target cells; therapeutic agents acting on tumors; or other therapeutically effective compounds for treating cancer or other diseases.

When used in combination with another chemotherapeutic agent, the compound of formula (I) can be formulated with one, two, three, four or more other therapeutic agents, for example, into a pharmaceutical form, which contains two, three, four, or more The therapeutic agent, that is, all the ingredients are contained in a single pharmaceutical composition. In another embodiment, the individual therapeutic agents can be formulated separately and presented together in a set, and optionally include instructions for their use.

In a specific embodiment, a compound of formula (I) is provided for use in combination with one or more (eg, 1 or 2) other therapeutic agents (eg, the above-mentioned anticancer agents). In yet another specific embodiment, a combination of an IAP antagonist as described herein and a PI3K / AKT pathway inhibitor selected from the group consisting of: apitolisib, buparisib, copanlisib, pictilisib (pictrelisib, ZSTK-474, CUDC-907, GSK-2636771, LY-3023414, ipatasertib, afuresertib, MK-2206, MK-8156, Idelalisib, BEZ235 (Datoris ( dactolisib)), BYL719, GDC-0980, GDC-0941, GDC-0032, and GDC-0068.

In another specific embodiment, a compound of formula (I) is provided in combination with one or more (eg, 1 or 2) other therapeutic agents (eg, anticancer agents) for use in treatment, such as prevention or treatment of cancer.

In a specific embodiment, the pharmaceutical composition comprises a compound of formula (I) and a pharmaceutically acceptable carrier, and optionally includes one or more therapeutic agents.

In another embodiment, the present invention relates to a combined use according to the present invention, which is used to make a pharmaceutical composition that inhibits tumor cell growth.

In another embodiment, the present invention relates to a product containing a compound of formula (I) and one or more anticancer agents, which is a combined preparation for simultaneous, separate or sequential use in the treatment of cancer patients on.

Examples

The invention will now be described with reference to specific embodiments described in the following examples, but is not limited to these examples. Compounds are named using an automated naming suite such as AutoNom (MDL), or as named by a chemical supplier.

The following synthetic steps are provided to illustrate the method used; for one of the preparations or steps, the precursors used are not necessarily derived from individual batches synthesized according to the steps in the description. In these examples, the following abbreviations are used.

NMR data: Unless otherwise stated, ¹ H NMR spectra were recorded on a Bruker Avance I spectrometer at 25 ° C and 400 MHz operation. Use Topspin 2.1 software to process and analyze data. In the NMR data, when the number of selected protons is less than the theoretical number of protons in the molecule, it is considered that the signal that obviously disappears is covered by the peak signal of the solvent and / or water. In addition, when the spectrum is obtained in a protonic NMR solvent, NH and / or OH protons exchange with the solvent, and such signals are generally not observed.

Analytical and Preparative LC-MS systems

Analytical LC-MS system and method description

In the following examples, compounds were characterized by a mass spectrometer using the following system and operating conditions. When atoms with different isotopes are present and reference a single mass, the compound reference mass is a monoisotopic mass (ie, ³⁵ Cl; ⁷⁹ Br, etc.).

Waters Platform LC-MS system: HPLC system: Waters 2795 Mass spectrometer: Micromass Platform LC PDA detector: Waters 2996 PDA Platform MS Conditions: Capillary voltage: 3.6 kV (3.40 kV at ES negative) cone ) Voltage: 30 V Source temperature: 120 ° C Scanning range: 125-800 amu Ionization method: ElectroSpray Positive or ElectroSpray Negative or Electrospray Positive and Negative

Waters Fractionlynx LC-MS System: HPLC System: 2767 Autosampler-2525 Binary Gradient Pump Mass Detector: Waters ZQ PDA Detector: Waters 2996 PDA Fractionlynx MS Conditions: Capillary Voltage: 3.5 kV (3.25 kV in ES (Cathode) cone voltage: 40 V (25 V on ES cathode) Source temperature: 120 ° C Scanning range: 125-800 amu Ionization method: electrospray positive or electrospray negative or electrospray positive and negative

Agilent 1200SL-6140 LC-MS system-RAPID: HPLC system: Agilent 1200 series SL mass detector: Agilent 6140 single quadrupole Second detector: Agilent 1200 MWD SL Agilent MS conditions: Capillary voltage: 4000V at ES anode (ES pos) (3500V at ES Neg) Fragmentor / Gain: 100 Gain: 1 Dry gas flow rate: 7.0 L / min Gas temperature: 345 ° C Nebuliser Pressure ): 35 psig Scanning range: 125-800 amu Ionization method: ElectroSpray Positive-Negative switching

Preparative LC-MS system and method description

Preparative LC-MS is a standard and effective method for the purification of small molecule organic compounds of compounds as described herein. The methods of liquid chromatography (LC) and mass spectrometry (MS) can be changed to provide better separation of crude products, and MS can improve sample detection. The preparative gradient LC method can be optimized by changing the chromatography column, volatile eluent and modifier, and gradient. These methods are known in the art to optimize preparative LC-MS, and compounds can be purified using these methods. Such methods are described in Rosentreter U, Huber U .; Optimal fraction collecting in preparative LC / MS; J Comb Chem . ; 2004; 6 (2), 159-64 and Leister W, Strauss K, Wisnoski D, Zhao Z, Lindsley C., Development of a custom high-throughput preparative liquid chromatography / mass spectrometer platform for the preparative purification and analytical analysis of compound libraries; J Comb Chem . ; 2003; 5 (3); 322-9.

Various systems for purifying compounds by preparative LC-MS are described below, however, those skilled in the art know that additional systems and methods can be used. From the information provided herein, or using other chromatography systems, those skilled in the art can purify the compounds described herein by preparative LC-MS.

Waters Fractionlynx system: Hardware: 2767 Dual Loop Autosampler / Fraction Collector 2525 preparative pump 2525 preparative pump (column fluid assembly for column selection) Column fluidic organiser) RMA (Waters reagent manager) as make up pump Waters ZQ mass spectrometer Waters 2996 Photo Diode Array detector Waters ZQ mass spectrometer Waters MS operating conditions: Capillary voltage: 3.5 kV (3.2 kV at ES cathode) cone voltage: 25 V Source temperature: 120 ° C Scanning range: 125-800 amu Ionization method: Electrospray positive (ElectroSpray Positive) or ElectroSpray Negative

Agilent 1100 LC-MS preparative system: Hardware: Autosampler: 1100 series “prepALS” Pump: 1100 series “PrepPump” for preparative flow gradient and pumping modifier in prep flow 1100 series "QuatPump" UV detector: 1100 series "MWD" multi-wavelength detector MS detector: 1100 series "LC-MSD VL" Dispensing collector: 2 x "Prep-FC" compensation pump ( Make Up pump): "Waters RMA" Agilent Active Splitter Agilent MS operating conditions: Capillary voltage: 4000 V (3500 V at ES cathode) Fragmentor / gain: 150/1 Dry gas flow rate: 12.0 L / min Gas temperature: 350 ° C Spray pressure: 50 psig Scanning range: 125-800 amu Ionization mode: ElectroSpray Positive or ElectroSpray Negative

Tubing:

Many commercially available columns, including non-palm and palm columns, can be used with different mobile phases, organic modifiers, and pH to achieve a wide range of selectivity. All tubing strings are used in accordance with the operating conditions recommended by the manufacturer. Generally speaking, a 5 micron particle size is used. For example, Waters (including, but not limited to, XBridge ^TM Prep OBD ^TM C18 and Phenyl, Atlantis® Prep T3 OBD ^TM and Sunfire ^TM Prep OBD C18 5 μm 19 x 100 mm), Phenomenex (including, but not limited to, Synergy MAX -RP and LUX ^TM Cellulose-2), Astec (Chirobiotic ^TM columns, including, but not limited to, V, V2, and T2) and Diacel® (including, but not limited to, Chiralpak® AD-H) For screening.

Extraction liquid:

Select the mobile phase eluent in conjunction with the stationary phase restrictions recommended by the column manufacturer to optimize column separation performance.

method:

Non-palm preparative chromatography

The compound examples were purified by HPLC, and the descriptions thereon were in accordance with the recommendations in the description of Snyder LR, Dolan JW, High-Performance Gradient Elution The Practical Application of the Linear-Solvent-Strength Model, Wiley, Hoboken, 2007. Method.

Palm preparative chromatography

Preparative separations using palm stationary phases (CSPs) are natural techniques using resolution of a mixture of isomers. In the same way, it can be applied to the separation of non-mirromeric isomers and non-palmable molecules. Methods for optimizing preparative palm separation on CSPs are well known in the art, and compounds are then purified using these methods. Such methods are described in Beesley T. E., Scott R.P.W .; Chiral Chromatography; Wiley, Chichester, 1998.

The values of salt stoichiometry or acid content in the compounds provided herein are experimentally obtained and will vary depending on the analytical method used. If the salt form is not specified, the resulting compound is in free base form.

Preparation Example 1: (R) -2-((S) -2-benzyloxycarbonylamino-3-hydroxy-propionyl-amino) -propionic acid methyl ester ((R) -2- ( (S) -2-Benzyloxycarbonylamino-3-hydroxy-propionyl-amino) -propionic acid methyl ester)

Diisopropylethylamine (375 mL) was dripped into (R) -2-amino-propionic acid methyl ester hydrochloride (100 g, 0.716 mol), EDC (165 g, 0.86 moles), carbobenzyloxy-L-serine (171.4 g, 0.716 moles) and DCM (3.6 L) in a cooled mixture. The resulting mixture was stirred under nitrogen and ambient temperature for 16 hours. After removal of the solvent in vacuo at 40 ^o C (in vacuo), the residue, water (1 L) was diluted with saturated sodium carbonate (1 L), and in EtOAc (2 L, 2 x 1 L) and extracted. The combined organic mixed with 2M hydrochloric acid (1 L), saturated brine solution (1L) washed, dried over magnesium sulfate, and concentrated at 40 ^o C in vacuo to give the title compound (172 g), as a colorless solid. ¹ H NMR (Me-d3-OD): 7.44-7.28 (6H, m), 5.13 (2H, s), 4.46 (1H, d), 4.43 (1H, d), 4.25 (1H, t), 3.82- 3.68 (5H, m), 1.39 (3H, d).

Preparation Example 2: (3S, 6R) -3-hydroxymethyl-6-methyl-piperazine-2,5-dione ((3S, 6R) -3-Hydroxymethyl-6-methyl-piperazine-2,5 -dione)

Under nitrogen at (R) -2-((S) -2-benzyloxycarbonylamino-3-hydroxy-propionyl-amino) -propionic acid methyl ester (which can be as described in Preparation Example 1 (Prepared as described above) (172 g, 0.53 moles) was added with 10% Pd / C (8.6 g), MeOH (530 mL) and cyclohexene (344 mL). The mixture was heated at reflux for 17 hours. MeOH (500 mL) was added and reflux was continued for 1 hour. The hot reaction mixture was filtered through a pad of celite and the cake was washed with hot MeOH (2 x 500 mL). The combined filtrate was concentrated. The obtained solid was slurried in 2-butanone (400 mL), and petrol (400 mL) was gradually added over a period of 10 minutes (over 10 min). After stirring for 30 min, the solid was filtered and the cake was washed with 2: 1 petrol / 2-butanone (300 mL). The filter cake was dried under vacuum at 40 ° C to obtain the title compound (68.3 g) as an off white solid. ¹ H NMR (DMSO-d6): 8.08 (1H, s), 7.90 (1H, s), 5.11 (1H, t), 3.92 (1H, q), 3.80-3.71 (1H, m), 3.71-3.60 ( 1H, m), 3.58-3.47 (1H, m), 1.24 (3H, d).

Preparation Example 3: ((2R, 5R) -5-methyl-piperazin-2-yl) -methanol hydrochloride (((2R, 5R) -5-methyl-piperazin-2-yl) -methanol hydrochloride )

(3S, 6R) -3-hydroxymethyl-6-methyl-piperazine-2,5-dione (which can be prepared as described in Preparation Example 2) (34 g, 0.215 moles) A solution of borane in THF (1 M, 1.6 L, 1.6 mol), and the mixture was heated to 70 ° C for 18 hours. The solution was cooled in an ice bath, and then MeOH (425 mL) was gradually added, followed by 5 M hydrochloric acid (113 mL). The mixture was heated to 70 ° C for 2 hours and then cooled to ambient temperature. The resulting solid was filtered, the cake was washed with THF (200 mL), and dried under vacuum at 40 ° C to give the title compound (39.3 g) as a colorless solid. ¹ H NMR (DMSO-d6): 9.79 (3H, s), 5.59 (1H, s), 3.76-3.40 (5H, m), 3.19-2.94 (2H, m), 1.28 (3H, d).

Preparation Example 4: (2R, 5R) -5-hydroxymethyl-2-methyl-piperazine-1-carboxylic acid third butyl ester ((2R, 5R) -5-Hydroxymethyl-2-methyl-piperazine-1 -carboxylic acid tert-butyl ester)

((2R, 5R) -5-methyl-piperazin-2-yl) -methanol hydrochloride in MeOH (96 mL) at 0 ° C (ice bath) (which can be used as in Preparation Example 3) (20 g, 119 mmol) was added to triethylamine (48.7 mL, 357 mmol). A tert-Butyl dicarbonate (61 g, 280 mmol) dissolved in MeOH (145 mL) was added over a period of 30 min. The reaction temperature was maintained at <10 ° C for 1 hour, and heated to ambient temperature over a period of 1 hour, and then heated to 50 ° C for 18 hours. The reaction solution was concentrated, and the residue was dissolved in ethanol (397 mL). A solution of NaOH (23.8 g, 595 mmol) dissolved in water (397 mL) was added, and the reaction was heated to 100 ° C for 18 hours, and then cooled to ambient temperature. The mixture was neutralized to pH 9 (using a pH meter) with 1M HCl (~ 300 mL), then extracted with chloroform (3 x 700 mL), dried over sodium sulfate, filtered, and concentrated. The residue was redissolved in MeOH and concentrated, then dried under vacuum at 40 ° C to give the title compound (21 g, 75%) as a colorless solid. ¹ H NMR (Me-d3-OD): 4.20-4.07 (1H, m), 3.79 (1H, dd), 3.71-3.58 (2H, m), 3.54 (1H, dd), 3.24 (1H, dd), 3.18-3.01 (1H, m), 3.01-2.89 (1H, m), 2.55 (1H, dd), 1.48 (9H, s), 1.25 (3H, s).

Preparation Example 5: (2R, 5R) -4-benzyl-5-hydroxymethyl-2-methyl-piperazine-1-carboxylic acid third butyl ester ((2R, 5R) -4-Benzyl-5 -hydroxymethyl-2-methyl-piperazine-1-carboxylic acid tert-butyl ester)

(2R, 5R) -5-hydroxymethyl-2-methyl-piperazine-1-carboxylic acid tert-butyl ester (which can be prepared as described in Preparation Example 4) (3.48 g, 15.1 mmol) , Benzaldehyde (1.76 g, 16.6 mmol), sodium triacetoxyborohydride (3.84 g, 18.1 mmol) and 1,2-dichloroethane (30 mL) in a mixture Stir at 20 ^o C for 18 hours and then partition it between saturated aqueous NaHCO ₃ (150 mL) and DCM (3 x 50 mL). The combined organic extracts were dried (Na ₂ SO ₄ ) and evaporated in vacuo to give an oil. Chromatography (SiO ₂ , 0-30% EtOAc / petrol) gave the title compound (4.588 g, 74%) as a colorless solid. MS: [M + H] ⁺ = 321.

Preparation Example 6: (2R, 5R) -4-benzyl-5-chloromethyl-2-methyl-piperazine-1-carboxylic acid third butyl ester ((2R, 5R) -4-Benzyl-5 -chloromethyl-2-methyl-piperazine-1-carboxylic acid tert-butyl ester)

Methanesulfonyl chloride (570 µL, 7.35 mmol) was added to (2R, 5R) -4-benzyl-5-hydroxymethyl with TEA (2.6 mL, 18.4 mmol) at 0 ° C. Of methyl-2-methyl-piperazine-1-carboxylic acid tert-butyl ester (which can be prepared as described in Preparation 5) (1.9 g, 6.12 mmol) in DCM (30 mL). The solution was stirred at room temperature for 18 hours. The reaction solution was partitioned between aqueous NH ₄ Cl and DCM. The organic phase was collected, dried over MgSO _4, filtered, and concentrated in vacuo. After chromatography _{(SiO 2, 30% EtOAc /} petrol), to give the title compound (1.6 g), as a white solid. MS: [M + H] ⁺ = 339.

Preparation Example 7: (2R, 5S) -4-benzyl-2-methyl-5-((R) -3-methyl-morpholin-4-ylmethyl) -piperazine-1-carboxylic acid Third butyl ester ((2R, 5S) -4-Benzyl-2-methyl-5-((R) -3-methyl-morpholin-4-ylmethyl) -piperazine-1-carboxylic acid tert-butyl ester)

K ₂ CO ₃ (81.6 g, 591 mmol) and KI (73.6 g, 443 mmol) were added to (2R, 5R) -4-benzyl-5-chloromethyl-2-methyl- A solution of piperazine-1-carboxylic acid third butyl ester (which can be prepared as described in Preparation Example 6) (50 g, 147.9 mmol) in acetonitrile (400 mL), followed by addition of (R) -3- Methyl-morpholine hydrochloride ((R) -3-methyl-morpholine hydrochloride) (26.4 g, 192 mmol). The reaction solvent was stirred at 70 ° C for 18 hours. The solid was removed by filtration, and the solvent was removed in vacuo. The crude product was purified by chromatography (using a pad of silica, 20% EtOAc / Petrol) to give the title compound (41.3 g) as a white solid. MS: [M + H] ⁺ = 404.

Preparation Example 8: (2R, 5S) -2-methyl-5-((R) -3-methyl-morpholin-4-ylmethyl) -piperazine-1-carboxylic acid third butyl ester (( 2R, 5S) -2-Methyl-5-((R) -3-methyl-morpholin-4-ylmethyl) -piperazine-1-carboxylic acid tert-butyl ester)

Add Pd / C (palladium on carbon) (10%) (33 g) and acetic acid (220 mL) to (2R, 5S) -4-benzyl-2-methyl-5-((R) -3 -Methyl-morpholin-4-ylmethyl) -piperazine-1-carboxylic acid tert-butyl ester (which can be prepared as described in Preparation Example 7) (41.3 g, 102 mmol) in EtOH (300 mL). The mixture was stirred for 18 hours at room temperature under H ₂ (one atmosphere). The reaction mixture was then filtered through a celite pad to remove the catalyst, and the solvent was removed in vacuo. The crude product was partitioned between saturated aqueous NaHCO ₃ and DCM, and to DCM (3x) and the product extracted. In the organic phase was dried MgSO _4, filtered, and concentrated in vacuo to give the title compound as a pale yellow (pale yellow) oil. ¹ H NMR (400 MHz, CDCl3): 4.43-3.87 (1H, m), 3.78 (1H, d), 3.73-3.55 (3H, m), 3.32 (1H, dd), 3.22 (1H, dd), 3.16 -2.93 (3H, m), 2.93-2.72 (1H, m), 2.55-2.35 (2H, m), 2.35-2.15 (2H, m), 1.89 (1H, dd), 1.45 (9H, s), 1.26 (3H, d), 0.96 (3H, d).

Another method:

Add (2R, 5S) -4-benzyl-2-methyl-5-((R) -3-methyl-morpholin-4-yl) to a sealed 10L flask with a stir bar. Methyl) -piperazine-1-carboxylic acid third butyl ester (500 g, 1.24 moles, 1.0 equivalent (eq)) (which can be prepared as described in Preparation Example 7) and ethanol (Stock, 5L). The flask was placed under nitrogen, and 10% Pd / C (Aldrich, 50 g, 0.124 mole, 0.1 equivalent) was added as a slurry in ethanol. The flask was purged several times with a di-vac pump and placed under a hydrogen atmosphere using 4 using a diaphragm vacuum pump (di-vac pump) balloons). The reaction was warmed to 30 ° C. overnight and it was confirmed by NMR that the starting material was completely consumed. The reaction mixture was cooled to room temperature and filtered through a pad of celite under nitrogen. The filtrate was evaporated to dryness to give the title product as a colorless oil.

¹ H NMR (MeOD): 1.00 (3H, d), 1.25 (3H, d), 1.48 (9H, s), 2.08-2.14 (1H, m), 2.28-2.35 (1H, m), 2.42-2.48 ( 1H, m), 2.49-2.55 (1H, dd), 2.80-3.06 (4H, m), 3.22-3.28 (2H, m), 3.61-3.78 (4H, m), 4.12-4.16 (1H, m).

¹³ C NMR (MeOD): 14.6, 15.7, 28.8, 40.8, 44.8, 48.3, 50.3, 53.2, 54.3, 57.5, 68.5, 73.9, 81.1, 157.0.

Preparation Example 9: (2R, 5S) -4-benzyl-5-((3R, 5R) -3,5-dimethyl-morpholin-4-ylmethyl) -2-methyl-piperazine Tert-butyl carboxylic acid ((2R, 5S) -4-Benzyl-5-((3R, 5R) -3,5-dimethyl-morpholin-4-ylmethyl) -2-methyl-piperazine-1- carboxylic acid tert-butyl ester)

K ₂ CO ₃ (2.7 g, 19.5 mmol) and KI (1.83 g, 11.05 mmol) were added to (2R, 5R) -4-benzyl-5-chloromethyl-2-methyl- Piperazine-1-carboxylic acid third butyl ester (which can be prepared as described in Preparation Example 6) (2.2 g, 6.5 mmol) in acetonitrile (30 mL), followed by (3R, 5R) -3 , 5-Dimethyl-morpholine (0.80 g, 7.0 mmol). The reaction was stirred at 70 ° C for 18 hours. Filter to remove solids and remove solvent in vacuo. The residue was partitioned between water and dichloromethane. The organic phase was dried, filtered, and the solvent was evaporated. The crude product was purified by silica chromatography (0-40% EtOAc in Petrol) to give the title compound (2.56 g, 94%) as a white solid. MS: [M + H] ⁺ = 418.

Preparation Example 10: (2R, 5S) -5-((3R, 5R) -3,5-dimethyl-morpholin-4-ylmethyl) -2-methyl-piperazine-1-carboxylic acid Tributyl ester ((2R, 5S) -5-((3R, 5R) -3,5-Dimethyl-morpholin-4-ylmethyl) -2-methyl-piperazine-1-carboxylic acid tert-butyl ester)

Add Pd / C (10%) (1.6 g) and acetic acid (10 mL) to (2R, 5S) -4-benzyl-5-((3R, 5R) -3,5-dimethyl-? Phenolin-4-ylmethyl) -2-methyl-piperazine-1-carboxylic acid tert-butyl ester (which can be prepared as described in Preparation Example 9) (2.5 g, 6.0 mmol) in EtOH (70 mL). The mixture was at room temperature under H ₂ (1 atm) was stirred for three hours. The reaction mixture was then filtered through a celite pad to remove the catalyst, and the solvent was removed in vacuo. The crude product was partitioned between saturated aqueous NaHCO ₃ and DCM, and to DCM (3x) and the product extracted. In the organic phase was dried MgSO _4, filtered, and concentrated in vacuo to give the title compound as a pale yellow (pale yellow) oil. ¹ H NMR (400 MHz, CDCl3): 4.16 (1H, s), 3.79-3.59 (3H, m), 3.44-3.19 (3H, m), 3.08 (1H, dd), 2.99-2.69 (4H, m) , 2.52 (1H, dd), 2.29 (1H, dd), 1.47 (9H, s), 1.27 (3H, d), 1.00 (6H, d).

The following compounds were prepared following similar steps to Preparations 9 and 10:

10A: (2R, 5S) -5-((2S, 5R) -2,5-dimethyl-morpholin-4-ylmethyl) -2-methyl-piperazine-1-carboxylic acid tert-butyl Ester ((2R, 5S) -5-((2S, 5R) -2,5-Dimethyl-morpholin-4-ylmethyl) -2-methyl-piperazine-1-carboxylic acid tert-butyl ester), MS: [M + H] ⁺ = 328.

Preparation Example 11: 2- (5-Chloro-3-fluoro-pyridin-2-yl) -2-methyl-propionitrile (2- (5-Chloro-3-fluoro-pyridin-2-yl) -2- methyl-propionitrile)

A solution of sodium bis (trimethylsilyl) amide (610 mL, 40% in tetrahydrofuran, 1.326 mol) was added to 5-chloro-2,3-difluoropyridine (198.2 g, 1.326 moles) and isobutyronitrile (238 mL, 2.65 moles) in toluene (2 L) in an ice-cold solution. The mixture was stirred at room temperature under nitrogen overnight, and then a saturated aqueous ammonium chloride solution (1 L) was added. The phases were separated and the aqueous layer was extracted with ethyl acetate (2 x 1 L). The combined organic extracts were dried (MgSO ₄₎ and concentrated in vacuo at 40 ^o C, to give the title compound (259.8 g, 95%). ¹ H NMR (400 MHz, DMSO-d6): 8.57 (1H, dd), 8.24 (1H, dd), 1.74 (6H, width).

Preparation Example 12: 2- (5-Chloro-3-fluoropyridin-2-yl) -2-methylpropylamine

Borane-tetrahydrofuran complex (1 M, 1.37 L, 1.365 moles) was added to 2- (5-chloro-3-fluoro-pyridin-2-yl) -2-methyl-propane Nitrile (which can be prepared as described in Preparation 11) (135.6 g, 0.683 moles) in a cold solution of tetrahydrofuran (670 mL). The mixture was stirred at room temperature under nitrogen overnight and then cooled in ice. The mixture was quenched by the addition of 5M hydrochloric acid (335 mL). The resulting mixture was basified with a 40% aqueous potassium hydroxide solution (460 mL), and the phases were separated. With ethyl acetate (2 x 670 mL) basic aqueous phase was extracted, and the combined organic extracts were washed with brine (670 mL) washed, dried (MgSO ₄₎ and concentrated in vacuo at 40 ^o C, to give the title compound (102.9 g , 74%). ¹ H NMR (400 MHz, DMSO-d6): 8.44 (1H, t), 7.95 (1H, dd), 2.85 (2H, d), 1.29 (6H, d).

Preparation Example 12, another step: 2- (5-chloro-3-fluoropyridin-2-yl) -2-methylpropylamine

In a 10 L flange flask, 2- (5-chloro-3-fluoro-pyridin-2-yl) -2-methyl-propionitrile (which can be prepared as described in Preparation Example 11) was added. (200 g, 1.00 mol), nickel (II) chloride hexahydrate (239.4 g, 1.00 mol), and ethanol (3.0 L). The resulting green solution was cooled to 0 ° C in a dry ice / acetone bath under a nitrogen atmosphere. Sodium borohydride (114.3 g, 3.02 mol) was added in portions at a rate such that the reaction temperature was maintained below 6 ° C (addition time = 1¾ hours) to obtain a black suspension. After the addition was complete, the cold bath was immediately replaced with an ice / water bath, and then the reaction was allowed to warm to room temperature overnight. The reaction mixture was cooled in an ice bath to 0 to 4 ° C. A 25% ammonia solution (2680 mL) was added using a dropping funnel, and the reaction temperature was maintained below 10 ° C (addition time = 1 hour). After the addition was complete, stirring was continued for 30 minutes at about 0 ° C. The mixture was then filtered through celite and the residue was washed with ethanol (2 x 750 mL). (Caution! Do not let the filter pad dry. The total filtration time is about 2 hours.) Transfer the pale yellow / brown filtrate to a large rotary evaporator and concentrate until all ethanol is removed. The resulting green oil was transferred to a 5 L separating funnel and a 25% ammonia solution was added until the oil turned yellow (200 mL). The oil was separated and the aqueous phase was extracted with toluene (2 x 300 mL). Wash the mixed organic extracts with 1: 1 25% ammonia solution / brine (300 mL), filter with sodium sulfate, and concentrate on a rotary evaporator (immersion bath temperature up to 70 ° C) to obtain the crude product as a yellow oil (161 g), the data is consistent with the above. This was used in the next step without purification.

Preparation Example 13: 6-chloro-3,3-dimethyl-2,3-dihydro-1H-pyrrolo [3,2-b] pyridine (6-chloro-3,3-dimethyl-2,3- dihydro-1H-pyrrolo [3,2-b] pyridine)

2- (5-chloro-3-fluoropyridin-2-yl) -2-methylpropylamine (which can be prepared as described in Preparation 12 and another step of Preparation 12) (33 g, 0.163 mol Ear), a mixture of potassium carbonate (122 g, 0.884 moles) and NMP (100 mL) was heated to 150 ^o C for 4 hours. The cooled mixture was diluted with water (330 mL) and extracted with toluene (3 x 300 mL). The combined organic extracts were washed with brine (160 mL) washed, dried (MgSO ₄₎ and concentrated in vacuo at 40 ^o C, to give a crude product (24.8 g). Silica chromatography, eluting with 5-30% ethyl acetate / petrol, gave the title compound (21 g, 71%). ¹ H NMR (400 MHz, DMSO-d6): 7.61 (1H, d), 6.75 (1H, d), 6.06 (1H, bs), 3.31 (2H, s), 1.21 (6H, s).

Preparation Example 14: 6-Chloro-3,3-dimethyl-2,3-dihydropyrrolo [3,2-b] pyridine-1-carboxylic acid tert-butyl ester (6-Chloro-3,3- dimethyl-2,3-dihydropyrrolo [3,2-b] pyridine-1-carboxylic acid tert-butyl ester)

Add a third butyl dicarbonate (3.7 g, 17.1 mmol) to 6-chloro-3,3-dimethyl-2,3-dihydro-1H-pyrrolo [3,2-b] pyridine (It can be prepared as described in Preparation Example 13) (2.6 g, 14.2 mmol), a mixture of tetrahydrofuran (26 mL) and 2 M sodium hydroxide (11.4 mL, 22.8 mmol), and continuously stirred over 2 day. This biphasic mixture was diluted with water (20 mL) and extracted with ethyl acetate (2 x 20 mL). The organic extract was mixed dried (MgSO ₄₎ and concentrated in vacuo at 40 ^o C, to give a crude product (6.02 g). Silica chromatography, eluting with 5-30% ethyl acetate / petrol, to give the title compound (2.23g, 55%); ¹ H NMR (400 MHz, DMSO-d6): 8.11 (1H, d) , 7.85 (1H, bs), 3.77 (2H, s), 1.52 (9H, s), 1.28 (6H, s).

Preparation Example 15: 6- (4-fluorobenzyl) -3,3-dimethyl-2,3-dihydro-1H-pyrrolo [3,2-b] pyridine (6- (4-Fluorobenzyl) -3,3-dimethyl-2,3-dihydro-1H-pyrrolo [3,2-b] pyridine)

4-fluorobenzyl zinc chloride (4-fluorobenzylzinc chloride) (2L of 0.5M THF solution, 1 mole) at 20 ^o C was added to 6-chloro-3,3-dimethyl-2, 3-dihydro-1H-pyrrolo [3,2-b] pyridine (which can be prepared as described in Preparation Example 13) (91.3 g, 0.5 mole), lithium bromide (130.3 g, 1.5 mole), and dichloride [1,3-bis (2,6-diisopropylphenyl) imidazol-2-ylidene] (3-chloropyridyl) palladium (II) ([1,3-bis (2,6-diisopropylphenyl ) imidazol-2-ylidene] (3-chloropyridyl) palladium (II) dichloride) (6.8 g, 0.01 mole) in a degassed mixture of THF (685 mL) and NMP (910 mL) and exothermic. The resulting dark mixture was stirred under nitrogen at room temperature for 18 hours. The reaction was quenched with 2.5% aqueous citric acid solution (900 mL) and extracted with toluene (2 x 900 mL). Mixing the organic phase with water (3 x 900 mL), brine (900 mL) washed, dried using MgSO _4, filtered and concentrated in vacuo. The resulting solid was slurried in petrol (450 mL) and toluene (100 mL). After stirring for 30 min, the solid was filtered and the filter cake was washed with petrol (2 x 90 mL). The filter cake was dried at 40 ^o C in vacuo to give the title compound (107.3 g), as a gray solid. ¹ H NMR (DMSO-d6): 7.60 (1H, d), 7.30-7.22 (2H, m), 7.15-7.06 (2H, m), 6.53 (1H, d), 5.64 (1H, s), 3.78 ( 2H, s), 3.22 (2H, d), 1.19 (6H, s).

The following compounds were prepared in a manner similar to that described in Preparation 15:

15A: 6-[(4-fluorophenyl) methyl] -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridine-1-carboxylic acid third butyl ester ( tert-Butyl 6-[(4-fluorophenyl) methyl] -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridine-1-carboxylate), MS: [M + H] ⁺ = 357.

15B: 6- (3-fluorobenzyl) -3,3-dimethyl-2,3-dihydro-1H-pyrrolo [3,2-b] pyridine, MS: [M + H] ⁺ = 257.

15C: 6-butyl-3,3-dimethyl-2,3-dihydro-1H-pyrrolo [3,2-b] pyridine, MS: [M + H] ⁺ = 205.

15D: 6- (2-fluorobenzyl) -3,3-dimethyl-2,3-dihydro-1H-pyrrolo [3,2-b] pyridine, MS: [M + H] ⁺ = 257.

15E: 6- (2,4-difluorobenzyl) -3,3-dimethyl-2,3-dihydro-1H-pyrrolo [3,2-b] pyridine

Preparation Example 16: 5-Bromo-6- (4-fluorobenzyl) -3,3-dimethyl-2,3-dihydro-1H-pyrrolo [3,2-b] pyridine (5-Bromo -6- (4-fluorobenzyl) -3,3-dimethyl-2,3-dihydro-1H-pyrrolo [3,2-b] pyridine)

6- (4-fluorobenzyl) -3,3-dimethyl-2,3-dihydro-1H-pyrrolo [3,2-b] pyridine (which can be prepared as described in Preparation 15) ) (88.5 g, 0.345 moles) in DMF (1.67 L) was cooled to -5 ^o C. Solid N-bromosuccinimide (61.5 g, 0.345 mol) was added in portions and exothermic. The mixture was stirred for 1 hour while warming to room temperature. Water (2.66 L) was added, exotherm, and the resulting mixture was stirred at room temperature for 18 hours. The solid was filtered and the filter cake was washed with water (270 mL). The filter cake was dissolved in THF (1.5 L), dried over MgSO ₄ , filtered, and concentrated in vacuo to give the title compound (109.7 g) as a yellow solid. ¹ H NMR (DMSO-d6): 7.29-7.20 (2H, m), 7.20-7.03 (2H, m), 6.64 (1H, s), 5.88 (1H, s), 3.89 (2H, s), 3.26 ( 2H, d), 1.20 (6H, s).

The following compounds were prepared in a manner similar to that of Preparation Example 16:

16A: 5-bromo-6- (3-fluorobenzyl) -3,3-dimethyl-2,3-dihydro-1H-pyrrolo [3,2-b] pyridine (5-Bromo-6 -(3-fluorobenzyl) -3,3-dimethyl-2,3-dihydro-1H-pyrrolo [3,2-b] pyridine), MS: [M + H] ⁺ = 335, 337.

16B: 5-bromo-6- (2-fluorobenzyl) -3,3-dimethyl-2,3-dihydro-1H-pyrrolo [3,2-b] pyridine, MS: [M + H] ⁺ = 335, 337.

16C: 5-bromo-6-butyl-3,3-dimethyl-2,3-dihydro-1H-pyrrolo [3,2-b] pyridine, MS: [M + H] ⁺ = 283, 285.

16D: 5-bromo-6- (2,4-difluorobenzyl) -3,3-dimethyl-2,3-dihydro-1H-pyrrolo [3,2-b] pyridine

Production Example 17: [6- (4-fluoro-benzyl) -3,3-dimethyl-2,3-dihydro-1H-pyrrolo [3,2-b] pyridin-5-yl]- Methanol ([6- (4-Fluoro-benzyl) -3,3-dimethyl-2,3-dihydro-1H-pyrrolo [3,2-b] pyridin-5-yl] -methanol)

5-bromo-6- (4-fluorobenzyl) -3,3-dimethyl-2,3-dihydro-1H-pyrrolo [3,2-b] pyridine cooled to -78 ° C (It can be prepared as described in Preparation 16) (22.8 g, 68.2 millimoles) in THF (300 mL) was added MeLi (1.6 M in Et ₂ O; 51.1 mL , 91.8 millimoles). Third butyl lithium (1.7 M in hexane; 96 Ml, 164 mmol) was then added over a period of 30 minutes. After 15 minutes, DMF (26 mL) was added and the mixture was stirred at -78 ° C for another 50 minutes. A saturated aqueous NH ₄ Cl (450 mL) solution was added, and the mixture was stirred at room temperature for 10 minutes. The organic layer was separated and the aqueous layer was extracted with EtOAc (2 x 150 mL). The organic portions were mixed together, washed with brine (200 mL), dried (MgSO ₄ ) and evaporated to give 6- (4-fluoro-benzyl) -3,3-dimethyl-2,3-dihydro -1H-pyrrolo [3,2-b] pyridine-5-carboxaldehyde (6- (4-fluoro-benzyl) -3,3-dimethyl-2,3-dihydro-1H-pyrrolo [3,2-b] pyridine-5-carbaldehyde), as a yellow solid, without further purification. MS: m / z = 285 (M + H ⁺ ) ⁺ . This product (~ 68 mmol) was suspended in MeOH (250 mL) and cooled in an ice bath. NaBH ₄ (3.4 g, 81.8 mmol) was added portion by portion over 5 minutes. The cooling device was removed and the mixture was stirred for another 20 minutes. The mixture was cooled in an ice bath, and then a 10% aqueous KHSO ₄ solution was carefully added over a period of 10 minutes (note: bubbling). After the mixture was stirred at room temperature (RT) for 5 minutes, it was re-cooled using an ice bath. The mixture was basified by the addition of a 50% aqueous NaOH solution (~ 18 mL), and then concentrated in vacuo to about one third volume. The resulting aqueous mixture was extracted with CH ₂ Cl ₂ (1 x 200 Ml, 2 x 100 mL), and the CH ₂ Cl ₂ layers were mixed together and dried (MgSO ₄ ). The CH ₂ Cl ₂ solution was concentrated under vacuum to about 30 mL, and then diluted with toluene (70 mL) to the product began to crystallize. Collected by filtration to give the product as a colorless crystalline solid (10.6 g). A second crop (2.1 g) was collected from the filtrate. The filtrate was concentrated and the remaining material was purified by SiO ₂ chromatography (eluted with 25-50% EtOAc / hexanes) to give a third crop (2.1 g); the title compound was obtained in a total yield of 14.8 g (76%, after 2 steps), MS: [M + H] ⁺ = 287.

Another step involves subsequent recrystallization using isopropanol.

The following compounds were prepared in a manner similar to that of Preparation 17:

(6-Butyl-3,3-dimethyl-2,3-dihydro-1H-pyrrolo [3,2-b] pyridin-5-yl) -methanol, MS: [M + H] ⁺ = 235.

Production Example 18: 2-chloro-1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [ 3,2-b] pyridin-1-yl} ethyl-1-one (2-Chloro-1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} ethan-1-one)

In [6- (4-fluoro-benzyl) -3,3-dimethyl-2,3-dihydro-1H-pyrrolo [3,2-b] pyridin-5-yl] -methanol (which Can be prepared as described in Preparation Example 17) (11.8 g, 41.3 mmol) in a cooled (~ 5 ° C) suspension of MeCN (175 mL) with chloroacetyl chloride (6.9 mL, 86.7 Mol). The cooling device was removed and the mixture was stirred at room temperature for 30 minutes. The mixture was then evaporated in vacuo and dissolved in MeOH (200 mL). K ₂ CO ₃ solution (12 g in 100 mL H ₂ O) was added, and the mixture was stirred at room temperature for 20 minutes, and then concentrated in vacuo to ~ one quarter volume. The aqueous mixture was extracted with CH ₂ Cl ₂ (1 x 100 mL, 2 x 30 mL), and the CH ₂ Cl ₂ layers were mixed together and dried (MgSO ₄ ). Evaporation in vacuo gave the product as a colorless crystalline solid (12.1 g, ~ 100%). MS: [M + H] ⁺ = 363.

The following compounds were prepared in a similar manner to that described in Preparation 18:

18A: 1- (2-chloroethylfluorenyl) -6-[(4-fluorophenyl) methyl] -3,3-dimethyl-1H, 2H, 3H, 4H, 5H-pyrrolo [3, 2-b] pyridin-5-one, MS: [M + H] ⁺ = 349.

18B: 2-chloro-1- {6-[(2-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3, 2-b] pyridin-1-yl} ethyl-1-one, MS: [M + H] ⁺ = 363.

18C: 1- (2-chloroethylfluorenyl) -6-[(4-fluorophenyl) methyl] -3,3,4-trimethyl-1H, 2H, 3H, 4H, 5H-pyrrolo [ 3,2-b] pyridin-5-one, MS: [M + H] ⁺ = 363.

18D: 1- (2-chloroethylfluorenyl) -6-[(2,4-fluorophenyl) methyl] -3,3-dimethyl-1H, 2H, 3H, 4H, 5H-pyrrolo [ 3,2-b] pyridin-5-one, MS: [M + H] ⁺ = 367.

18E: 1- (2-chloroethylfluorenyl) -6-[(2-fluorophenyl) methyl] -3,3,4-trimethyl-1H, 2H, 3H, 4H, 5H-pyrrolo [ 3,2-b] pyridin-5-one, MS: [M + H] ⁺ = 363.

18F: 2-chloro-1- {6-[(2,4-difluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} ethan-1-one, MS: [M + H] ⁺ = 381.

18G: 2-chloro-1- [5- (1,2-dihydroxyethyl) -6-[(4-fluorophenyl) methyl] -3,3-dimethyl-1H, 2H, 3H-pyrrole And [3,2-b] pyridin-1-yl] ethan-1-one, MS: [M + H] ⁺ = 393.

18H: 2-chloro-1- {6-[(3-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3, 2-b] pyridin-1-yl} ethyl-1-one, MS: [M + H] ⁺ = 363.

18I: 1- (2-chloro-ethylamyl) -6- (4-fluoro-benzyl) -3,3-dimethyl-1,2,3,6-tetrahydro-pyrrolo [2, 3-c] pyridin-5-one, MS: [M + H] ⁺ = 349.

18J: 1- (2-chloro-ethylfluorenyl) -6- (2,4-difluoro-benzyl) -3,3-dimethyl-1,2,3,6-tetrahydro-pyrrolo [2,3-c] pyridin-5-one, MS: [M + H] ⁺ = 367.

18K: 2-chloro-1- [5-((R or S) -1,2-dihydroxyethyl) -6-[(4-fluorophenyl) methyl] -3,3-dimethyl- 1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl] ethan-1-one is obtained from the slower eluting precursor. MS: [M + H] ⁺ = 393.

18L: 1- (2-chloroethylfluorenyl) -6-[(2,4-fluorophenyl) methyl] -3,3,4-trimethyl-1H, 2H, 3H, 4H, 5H-pyrrole [3,2-b] pyridin-5-one, MS: [M + H] ⁺ = 381.

18M: 2-chloro-1- {6-[(4-fluorophenyl) methyl] -5- (R or S) -1-hydroxy-2-methoxyethyl) -3,3-dimethyl -1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} ethan-1-one, obtained from faster eluting precursor, MS: [M + H] ⁺ = 407.

18N: 2-chloro-1- {6-[(4-fluorophenyl) methyl] -5- (R or S) -1-methoxy-2-hydroxyethyl) -3,3-dimethyl -1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} ethan-1-one, from a faster elution precursor, MS: [M + H] ⁺ = 407.

18O: 2-chloro-1- {6-[(4-fluorophenyl) methyl] -5- (R or S) -1-hydroxy-2-methoxyethyl) -3,3-dimethyl -1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} ethan-1-one, from a slower elution precursor, MS: [M + H] ⁺ = 407.

18P: 2-chloro-1- {6-[(4-fluorophenyl) methyl] -5- (R or S) -1-methoxy-2-hydroxyethyl) -3,3-dimethyl -1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} ethan-1-one, from a slower elution precursor, MS: [M + H] ⁺ = 407.

18Q: 1- (2-chloro-ethenyl) -6- (2,4-difluoro-benzyl) -3,3,4-trimethyl-1,2,3,6-tetrahydro- Pyrrolop [2,3-c] pyridin-5-one, MS: [M + H] ⁺ = 381.

18R: 6-butyl-1- (2-chloro-ethylfluorenyl) -3,3-dimethyl-1,2,3,4-tetrahydro-pyrrolo [3,2-b] pyridine-5 -Ketone, MS: [M + H] ⁺ = 297.

18S: 1- [6-butyl-5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl] -2- Chloroethyl-1-one, MS: [M + H] ⁺ = 311.

18T: 6-butyl-1- (2-chloro-ethylfluorenyl) -3,3-dimethyl-1,2,3,6-tetrahydro-pyrrolo [2,3-c] pyridine-5 -Ketone, MS: [M + H] ⁺ = 297.

Preparation Example 19: (2R, 5S) -4- (2- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-oxoethyl) -2-methyl-5-{[((3R) -3-methylmorpholin-4-yl ] Methyl} piperazine-1-carboxylic acid tert-butyl ester (tert-Butyl (2R, 5S) -4- (2- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3, 3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-oxoethyl) -2-methyl-5-{[(3R) -3-methylmorpholin-4-yl] methyl} piperazine-1-carboxylate)

(2R, 5S) -2-methyl-5-((R) -3-methyl-morpholin-4-ylmethyl) -piperazine-1-carboxylic acid tert-butyl ester (which can be prepared as (Prepared as described in Example 8) (15.5 g, 46.4 mmol), KI (12.8 g, 77.4 mmol) and K ₂ CO ₃ (21.4 g, 155 mmol) were stirred in MeCN (70 mL), And cooled in an ice bath. Then 2-chloro-1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3, 2-b] pyridin-1-yl} ethan-1-one (which can be prepared as described in Preparation 18) (14.0 g, 38.7 mmol) in MeCN (100 mL). The mixture was stirred at room temperature for 2 hours, and then concentrated in vacuo to about a quarter volume. The mixture was partitioned between EtOAc (150 mL) and H ₂ O (150 mL), and the aqueous layer was extracted with EtOAc (1 x 75 mL). Together with the 10% of KH ₂ PO ₄ aqueous layers were combined EtOAc (4 x 100 mL), then brine (70 mL) wash. The organic layer was dried (MgSO _4), and evaporated to give the product as a colorless solid (25.8 g, 98%), MS: [M + H] + = 640.

The following compounds were prepared in a similar manner to that of Preparation Example 19:

(2R, 5S) -5-{[((3R, 5R) -3,5-dimethylmorpholin-4-yl] methyl) -4- (2- {6-[(4-fluorophenyl) (Methyl) -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-oxoethyl) Tert-Butyl (2R, 5S) -5-{[(3R, 5R) -3,5-dimethylmorpholin-4-yl] methyl} -4 -(2- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2 -oxoethyl) -2-methylpiperazine-1-carboxylate), MS: [M + H] ⁺ = 654.

(2R, 5S) -5-{[((3R, 5R) -3,5-dimethylmorpholin-4-yl] methyl) -4- (2- {6-[(4-fluorophenyl) Methyl] -3,3-dimethyl-5- pendantoxy-1H, 2H, 3H, 4H, 5H-pyrrolo [3,2-b] pyridin-1-yl} -2- pendantoxyethyl (Methyl) -2-methylpiperazine-1-carboxylic acid tert-butyl ester, MS: [M + H] ⁺ = 640.

(2R, 5S) -4- (2- {6-[(2-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-oxoethyl) -2-methyl-5-{[((3R) -3-methylmorpholin-4-yl] methyl} Piperazine-1-carboxylic acid tert-butyl ester, MS: [M + H] ⁺ = 640.

(2R, 5S) -4- (2- {6-[(4-fluorophenyl) methyl] -3,3-dimethyl-5- pendantoxy-1H, 2H, 3H, 4H, 5H- Pyrrolo [3,2-b] pyridin-1-yl} -2-oxoethyl) -2-methyl-5-{[((3R) -3-methylmorpholin-4-yl] methyl } Piperazine-1-carboxylic acid tert-butyl ester, MS: [M + H] ⁺ = 626.

(2R, 5S) -4- (2- {6-[(4-fluorophenyl) methyl] -3,3,4-trimethyl-5- pendant oxy-1H, 2H, 3H, 4H, 5H-pyrrolo [3,2-b] pyridin-1-yl} -2-oxoethyl) -2-methyl-5-{[((3R) -3-methylmorpholin-4-yl ] Methyl} piperazine-1-carboxylic acid tert-butyl ester, MS: [M + H] ⁺ = 640.

(2R, 5S) -4- (2- {6-[(2,4-difluorophenyl) methyl] -3,3-dimethyl-5- pendantoxy-1H, 2H, 3H, 4H , 5H-pyrrolo [3,2-b] pyridin-1-yl} -2-oxoethyl) -5-{[(3R, 5R) -3,5-dimethylmorpholine-4- Propyl] methyl} -2-methylpiperazine-1-carboxylic acid tert-butyl ester, MS: [M + H] ⁺ = 658.

(2R, 5S) -4- (2- {6-[(2,4-difluorophenyl) methyl] -3,3-dimethyl-5- pendantoxy-1H, 2H, 3H, 4H , 5H-pyrrolo [3,2-b] pyridin-1-yl} -2-oxoethyl) -2-methyl-5-{[((3R) -3-methylmorpholine-4- [Methyl] methyl} piperazine-1-carboxylic acid tert-butyl ester, MS: [M + H] ⁺ = 644.

(2R, 5S) -4- (2- {6-[(2-fluorophenyl) methyl] -3,3,4-trimethyl-5- pendantoxy-1H, 2H, 3H, 4H, 5H-pyrrolo [3,2-b] pyridin-1-yl} -2-oxoethyl) -2-methyl-5-{[((3R) -3-methylmorpholin-4-yl ] Methyl} piperazine-1-carboxylic acid tert-butyl ester, MS: [M + H] ⁺ = 640.

(2R, 5S) -5-{[((3R, 5R) -3,5-dimethylmorpholin-4-yl] methyl) -4- (2- {6-[(4-fluorophenyl) Methyl] -3,3,4-trimethyl-5- pendantoxy-1H, 2H, 3H, 4H, 5H-pyrrolo [3,2-b] pyridin-1-yl} -2-oxo Methyl ethyl) -2-methylpiperazine-1-carboxylic acid tert-butyl ester, MS: [M + H] ⁺ = 654.

(2R, 5S) -4- (2- {6-[(2,4-difluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H -Pyrrolo [3,2-b] pyridin-1-yl} -2-oxoethyl) -2-methyl-5-{[((3R) -3-methylmorpholin-4-yl] Methyl} piperazine-1-carboxylic acid tert-butyl ester, 1H NMR (400 MHz, Me-d3-OD): 8.12 (1H, s), 7.27-7.16 (1H, m), 7.06-6.86 (2H, m), 4.76 (2H, s), 4.17 (1H, s), 4.10-4.07 (2H, m), 3.99 (1H, d), 3.74-3.49 (5H, m), 3.30-3.22 (2H, m) , 2.97-2.77 (4H, m), 2.59-2.43 (2H, m), 2.43-2.32 (1H, m), 2.32-2.21 (1H, m), 1.47 (9H, s), 1.43 (6H, s) , 1.22 (3H, d), 1.00 (3H, d).

(2R, 5S) -4- {2- [5- (1,2-dihydroxyethyl) -6-[(4-fluorophenyl) methyl] -3,3-dimethyl-1H, 2H , 3H-pyrrolo [3,2-b] pyridin-1-yl] -2-oxoethyl} -2-methyl-5-{[((3R) -3-methylmorpholine-4- [Methyl] methyl} piperazine-1-carboxylic acid tert-butyl ester, MS: [M + H] ⁺ = 670; chiral HPLC (heptane / ethanol, 80:20, 0.2% DEA, chiralPAk -IC column) to obtain the faster eluting non-image isomer A, MS: [M + H] ⁺ = 670 and the slower eluting non-image isomer B, MS: [M + H] ⁺ = 670.

(2R, 5S) -4- (2- {6-[(3-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-oxoethyl) -2-methyl-5-{[((3R) -3-methylmorpholin-4-yl] methyl} Piperazine-1-carboxylic acid tert-butyl ester, MS: [M + H] ⁺ = 640.

(2R, 5S) -5-{[((3R, 5R) -3,5-dimethylmorpholin-4-yl] methyl) -4- (2- {6-[(4-fluorophenyl) Methyl] -3,3-dimethyl-5- pendantoxy-1H, 2H, 3H, 5H, 6H-pyrrolo [2,3-c] pyridin-1-yl} -2- pendantoxyethyl (Methyl) -2-methylpiperazine-1-carboxylic acid tert-butyl ester, MS: [M + H] ⁺ = 640.

(2R, 5S) -4- (2- {6-[(4-fluorophenyl) methyl] -3,3-dimethyl-5- pendant oxy-1H, 2H, 3H, 5H, 6H- Pyrrolo [2,3-c] pyridin-1-yl} -2-oxoethyl) -2-methyl-5-{[((3R) -3-methylmorpholin-4-yl] methyl } Piperazine-1-carboxylic acid tert-butyl ester, MS: [M + H] ⁺ = 626.

(2R, 5S) -5-{[((2S, 5R) -2,5-dimethylmorpholin-4-yl] methyl) -4- (2- {6-[(4-fluorophenyl) Methyl] -3,3,4-trimethyl-5- pendantoxy-1H, 2H, 3H, 4H, 5H-pyrrolo [3,2-b] pyridin-1-yl} -2-oxo Methyl ethyl) -2-methylpiperazine-1-carboxylic acid tert-butyl ester, MS: [M + H] ⁺ = 654.

(2R, 5S) -4- (2- {6-[(2,4-difluorophenyl) methyl] -3,3-dimethyl-5- pendantoxy-1H, 2H, 3H, 5H , 6H-pyrrolo [2,3-c] pyridin-1-yl} -2-oxooxyethyl) -5-{[((3R, 5R) -3,5-dimethylmorpholine-4- Propyl] methyl} -2-methylpiperazine-1-carboxylic acid tert-butyl ester, MS: [M + H] ⁺ = 658.

(2R, 5S) -4- {2- [5-((R or S) -1,2-dihydroxyethyl) -6-[(4-fluorophenyl) methyl] -3,3-di Methyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl] -2-oxoethylethyl} -5-{[(3R, 5R) -3,5-dimethyl Morpholine-4-yl] methyl} -2-methylpiperazine-1-carboxylic acid tert-butyl ester, MS: [M + H] ⁺ = 684.

(2R, 5S) -4- (2- {6-[(2,4-difluorophenyl) methyl] -3,3-dimethyl-5- pendantoxy-1H, 2H, 3H, 5H , 6H-pyrrolo [2,3-c] pyridin-1-yl} -2-oxoethyl) -5-{[(2S, 5R) -2,5-dimethylmorpholine-4- Propyl] methyl} -2-methylpiperazine-1-carboxylic acid tert-butyl ester, MS: [M + H] ⁺ = 658.

(2R, 5S) -4- (2- {4-amino-6-[(2,4-difluorophenyl) methyl] -3,3-dimethyl-5- pendantoxy-1H, 2H, 3H, 4H, 5H-pyrrolo [3,2-b] pyridin-1-yl} -2-oxoethyl) -2-methyl-5-{[((3R) -3-methyl Morpholine-4-yl] methyl} piperazine-1-carboxylic acid tert-butyl ester, MS: [M + H] ⁺ = 659.

(2R, 5S) -4- (2- {4-Amino-6-[(4-fluorophenyl) methyl] -3,3-dimethyl-5- pendantoxy-1H, 2H, 3H , 4H, 5H-pyrrolo [3,2-b] pyridin-1-yl} -2-oxoethyl) -2-methyl-5-{[((3R) -3-methylmorpholine- 4-yl] methyl} piperazine-1-carboxylic acid tert-butyl ester, MS: [M + H] ⁺ = 641.

(2R, 5S) -4- (2- {6-[(2,4-difluorophenyl) methyl] -3,3,4-trimethyl-5- pendantoxy-1H, 2H, 3H , 4H, 5H-pyrrolo [3,2-b] pyridin-1-yl} -2-sideoxyethyl) -5-{[(2S, 5R) -2,5-dimethylmorpholine- 4-yl] methyl} -2-methylpiperazine-1-carboxylic acid tert-butyl ester, MS: [M + H] ⁺ = 672.

(2R, 5S) -4- (2- {6-[(4-fluorophenyl) methyl] -5-((R or S) 1-hydroxy-2-methoxyethyl) -3,3- Dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-oxoethyl) -2-methyl-5-{[(3R) -3- Methylmorpholin-4-yl] methyl} piperazine-1-carboxylic acid third butyl ester (from faster eluting isomer), MS: [M + H] ⁺ = 684.

(2R, 5S) -4- (2- {6-[(4-fluorophenyl) methyl] -5-((R or S) 1-hydroxy-2-methoxyethyl) -3,3- Dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-oxoethyl) -2-methyl-5-{[(3R) -3- Methylmorpholine-4-yl] methyl} piperazine-1-carboxylic acid tert-butyl ester (from slower eluting isomer), MS: [M + H] ⁺ = 684.

(2R, 5S) -4- (2- {6-[(4-fluorophenyl) methyl] -5-((R or S) 1-methoxy-2-hydroxyethyl) -3,3 -Dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-oxoethyl) -2-methyl-5-{[(3R) -3 -Methylmorpholin-4-yl] methyl} piperazine-1-carboxylic acid tert-butyl ester (from the faster eluting isomer), MS: [M + H] ⁺ = 684.

(2R, 5S) -4- (2- {6-[(4-fluorophenyl) methyl] -5-((R or S) 1-methoxy-2-hydroxyethyl) -3,3 -Dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-oxoethyl) -2-methyl-5-{[(3R) -3 -Methylmorpholin-4-yl] methyl} piperazine-1-carboxylic acid tert-butyl ester (from the slower isomer), MS: [M + H] ⁺ = 684.

(2R, 5S) -4- (2- {4-Amino-6-butyl-3,3-dimethyl-5- pendantoxy-1H, 2H, 3H, 4H, 5H-pyrrolo [3 , 2-b] pyridin-1-yl} -2-oxoethyl) -5-{[((3R, 5R) -3,5-dimethylmorpholin-4-yl] methyl} -2 -Methylpiperazine-1-carboxylic acid tert-butyl ester, MS: [M + H] ⁺ = 603.

(2R, 5S) -4- (2- {6-[(2,4-difluorophenyl) methyl] -3,3,4-trimethyl-5- pendantoxy-1H, 2H, 3H , 5H, 6H-pyrrolo [2,3-c] pyridin-1-yl} -2-oxoethyl) -2-methyl-5-{[((3R) -3-methylmorpholine- 4-yl] methyl} piperazine-1-carboxylic acid tert-butyl ester, MS: [M + H] ⁺ = 658.

(2R, 5S) -4- (2- {6-butyl-3,3-dimethyl-5- pendantoxy-1H, 2H, 3H, 4H, 5H-pyrrolo [3,2-b] Pyridine-1-yl} -2-oxoethyl) -5-{[((3R, 5R) -3,5-dimethylmorpholin-4-yl] methyl} -2-methylpiperazine Tert-butyl-1-carboxylic acid, MS: [M + H] ⁺ = 588.

(2R, 5S) -4- (2- {6-butyl-3,3,4-trimethyl-5- pendantoxy-1H, 2H, 3H, 4H, 5H-pyrrolo [3,2- b] pyridin-1-yl} -2-oxoethyl) -5-{[((3R, 5R) -3,5-dimethylmorpholin-4-yl] methyl} -2-methyl Piperazine-1-carboxylic acid tert-butyl ester, MS: [M + H] ⁺ = 602.

(2R, 5S) -4- {2- [6-butyl-5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridine- 1-yl] -2-oxoethylethyl} -5-{[((3R, 5R) -3,5-dimethylmorpholin-4-yl] methyl} -2-methylpiperazine-1 -Tert-butyl carboxylic acid, MS: [M + H] ⁺ = 602.

(2R, 5S) -4- (2- {6-butyl-3,3-dimethyl-5- pendantoxy-1H, 2H, 3H, 5H, 6H-pyrrolo [2,3-c] Pyridine-1-yl} -2-oxoethyl) -5-{[((3R, 5R) -3,5-dimethylmorpholin-4-yl] methyl} -2-methylpiperazine Tert-butyl-1-carboxylic acid, MS: [M + H] ⁺ = 588.

(2R, 5S) -5-{[((3R, 5R) -3,5-dimethylmorpholin-4-yl] methyl) -4- (2- {6-[(4-fluorophenyl) Methyl] -5-((R or S) -2-hydroxy-1-methoxyethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridine- 1-yl} -2-oxooxyethyl) -2-methylpiperazine-1-carboxylic acid tert-butyl ester, MS: [M + H] ⁺ = 698.

(2R, 5S) -4- (2- {6-butyl-3,3-dimethyl-5- pendantoxy-1H, 2H, 3H, 4H, 5H-pyrrolo [3,2-b] Pyridin-1-yl} -2-oxoethyl) -5-{[((2S, 5R) -2,5-dimethylmorpholin-4-yl] methyl} -2-methylpiperazine Tert-butyl-1-carboxylic acid, MS: [M + H] ⁺ = 588.

Preparation Example 20: (2R, 5S) -4- {2- [6- (2,4-difluoro-benzyl) -3,3,4-trimethyl-5- pendant oxygen-2,3 , 4,5-tetrahydro-pyrrolo [3,2-b] pyridin-1-yl] -2-oxo-ethyl} -5-((3R, 5R) -3,5-dimethyl -Morpholine-4-ylmethyl) -2-methyl-piperazine-1-carboxylic acid tert-butyl ester ((2R, 5S) -4- {2- [6- (2,4-Difluoro-benzyl ) -3,3,4-trimethyl-5-oxo-2,3,4,5-tetrahydro-pyrrolo [3,2-b] pyridin-1-yl] -2-oxo-ethyl} -5-(( 3R, 5R) -3,5-dimethyl-morpholin-4-ylmethyl) -2-methyl-piperazine-1-carboxylic acid tert-butyl ester)

Add (2R, 5S) -4- {2- [6- (2,4-difluoro-benzyl) -3,3-dimethyl-5- pendantoxy-2,3,4,5- Tetrahydro-pyrrolo [3,2-b] pyridin-1-yl] -2-oxo-ethyl} -5-((3R, 5R) -3,5-dimethyl-morpholine-4 -Methyl) -2-methyl-piperazine-1-carboxylic acid tert-butyl ester (146 mg, 0.22 mmol) was dissolved in DMF (3 mL). Sodium hydride (60%, 11 mg, 0.27 mmol) was added and the mixture was stirred for 30 minutes. Methyl iodide (0.017 mL, 0.27 mmol) was added, and the mixture was stirred at room temperature for 30 minutes, then partitioned between water (10 mL) and EtOAc (2 x 10 mL). The organic portion was washed with brine, dried over magnesium sulfate, and concentrated. The residue was subjected to column chromatography, which was eluted with an eluent of 0-10% MeOH in EtOAc, and then purified by preparative HPLC to give the title compound (17.6 mg). MS: [M + H] ⁺ = 672.

Preparation Example 21: (2R, 5S) -4- {2- [6- (2,4-difluoro-benzyl) -3,3,4-trimethyl-5- pendantoxy-2,3 , 4,5-tetrahydro-pyrrolo [3,2-b] pyridin-1-yl] -2-oxo-ethyl} -2-methyl-5-((R) -3-methyl -Morpholin-4-ylmethyl) -piperazine-1-carboxylic acid tert-butyl ester ((2R, 5S) -4- {2- [6- (2,4-Difluoro-benzyl) -3,3 , 4-trimethyl-5-oxo-2,3,4,5-tetrahydro-pyrrolo [3,2-b] pyridin-1-yl] -2-oxo-ethyl} -2-methyl-5-((R ) -3-methyl-morpholin-4-ylmethyl) -piperazine-1-carboxylic acid tert-butyl ester)

Add (2R, 5S) -4- {2- [6- (2,4-difluoro-benzyl) -3,3-dimethyl-5- pendantoxy-2,3,4,5- Tetrahydro-pyrrolo [3,2-b] pyridin-1-yl] -2-oxo-ethyl} -2-methyl-5-((R) -3-methyl-morpholine-4 -Methyl) -piperazine-1-carboxylic acid tert-butyl ester (670 mg, 1.04 mmol) was dissolved in THF (20 mL). Lithium tert-butoxide (170 mg, 2.08 mmol) was added, followed by methyl iodide (0.16 mL, 2.60 mmol). The reaction was stirred at room temperature overnight and then partitioned between water (30 mL) and EtOAc (2 x 30 mL). The organic portion was washed with brine, dried over magnesium sulfate, and concentrated. The residue was purified by column chromatography, which was eluted with an eluent of 0-10% MeOH in DCM to give the title compound (350 mg). MS: [M + H] ⁺ = 658.

The following compounds were prepared in a similar manner to that of Preparation Example 21:

21A: (2R, 5S) -4- [2- (6-butyl-3,3,4-trimethyl-5- pendantoxy-2,3,4,5-tetrahydro-pyrrolo [3 , 2-b] pyridin-1-yl) -2-oxo-ethyl] -5-((3R, 5R) -3,5-dimethyl-morpholin-4-ylmethyl) -2 -Methyl-piperazine-1-carboxylic acid tert-butyl ester, MS: [M + H] ⁺ = 602.

Preparation Example 22: 6-[(4-fluorophenyl) methyl] -3,3-dimethyl-4-oxy-1H, 2H, 3H-pyrrolo [3,2-b] pyridine-1- Tert-Butyl 6-[(4-fluorophenyl) methyl] -3,3-dimethyl-4-oxy-1H, 2H, 3H-pyrrolo [3,2-b] pyridine-1-carboxylate )

6-[(4-fluorophenyl) methyl] -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridine-1-carboxylic acid at ambient temperature Butyl ester (which can be prepared as described in Preparation 15A) (3.88 g, 10.9 mmol) in a stirred solution of DCM (30 mL), and 3-chloroperbenzoic acid was added portionwise over a period of 0.1 hour. (chloroperbenzoic acid) (77%, 2.7 g, 12.0 mmol). The mixture was stirred for 3 hours and then partitioned between DCM (3 x 30 mL) and saturated aqueous NaHCO ₃ (150 mL). The organic extracts were mixed together, dried (Na ₂ SO ₄₎ and evaporated in vacuo. The residue was crystallized from ether-petrol to obtain the title compound (2.62 g). 1H NMR (400 MHz, Me-d3-OD): 7.74 (1H, s), 7.35-7.24 (2H, m), 7.13-7.02 (2H, m), 3.96 (2H, s), 3.79 (2H, s ), 1.57 (6H, s), 1.53 (9H, s).

Preparation Example 23: 6-[(4-fluorophenyl) methyl] -3,3-dimethyl-5- pendantoxy-1H, 2H, 3H, 4H, 5H-pyrrolo [3,2-b ] Pyridine-1-carboxylic acid tert-butyl ester (tert-Butyl 6-[(4-fluorophenyl) methyl] -3,3-dimethyl-5-oxo-1H, 2H, 3H, 4H, 5H-pyrrolo [3, 2-b] pyridine-1-carboxylate)

Heat 6-[(4-fluorophenyl) methyl] -3,3-dimethyl-4-oxy-1H, 2H, 3H-pyrrolo [3,2-b] pyridine-1 at 105 ^o C -A mixture of tert-butyl carboxylic acid (which can be prepared as described in Preparation 22) (0.6 g, 1.6 mmol) and acetic anhydride (4 mL) for 2 hours, then at 140 ^o C For 3 hours, cool, then pour the resulting solution into ice-water (~ 100 g). The resulting colorless solid was collected by filtration and suspended in methanol (15 mL). Aqueous NaOH (1 M, 1.8 mL) was added, and the mixture was stirred for 0.25 hours. The solution was concentrated in vacuo to 12 mL, then diluted with water (20 mL), and the resulting solid was collected by filtration to give the title compound (0.6 g). MS: [M + H] ⁺ = 373.

The following compounds were prepared in a manner similar to that of Preparation 23:

23A: 6-[(2-fluorophenyl) methyl] -3,3-dimethyl-5- pendantoxy-1H, 2H, 3H, 4H, 5H-pyrrolo [3,2-b] pyridine Tert-butyl carboxylic acid

Preparation Example 24: 6-[(4-fluorophenyl) methyl] -3,3-dimethyl-5- pendantoxy-1H, 2H, 3H, 4H, 5H-pyrrolo [3,2-b ] Pyridine (6-[(4-Fluorophenyl) methyl] -3,3-dimethyl-5-oxo-1H, 2H, 3H, 4H, 5H-pyrrolo [3,2-b] pyridine)

6-[(4-fluorophenyl) methyl] -3,3-dimethyl-5- pendantoxy-1H, 2H, 3H, 4H, 5H-pyrrolo [3,2-b] pyridine- A mixture of 1-carboxylic acid third butyl ester (which can be prepared as described in Preparation Example 23) (0.6 g, 1.6 mmol), methanol (20 mL) and 5 M aqueous HCl (20 mL) was heated and refluxed (reflux) for 16 hours, cooled and then treated with water. The obtained solid was collected by filtration to obtain the title compound (0.255 g). MS: [M + H] ⁺ = 273.

Preparation Example 25: 1- [5-Bromo-6- (3-fluoro-benzyl) -3,3-dimethyl-2,3-dihydro-pyrrolo [3,2-b] pyridine-1 -Yl] -ethyl ketone (1- [5-Bromo-6- (3-fluoro-benzyl) -3,3-dimethyl-2,3-dihydro-pyrrolo [3,2-b] pyridin-1-yl] -ethanone)

5-Bromo-6- (3-fluoro-benzyl) -3,3-dimethyl-2,3-dihydro-1H-pyrrolo [3,2-b] pyridine (which can be used as in the preparation example (Prepared as described in 16A) (4.5 g, 13.43 mmol) in toluene (50 mL) was added with acetyl chloride (1.05 mL, 14.78 mmol), and the reaction mixture was stirred at room temperature Overnight. Saturated aqueous NaHCO ₃ (50 mL), and to EtOAc (2x40 mL) and the product extracted. The organic phase was washed with brine, dried, filtered, and the solvent was evaporated to give the title compound (4.99 g). MS: [M + H] ⁺ = 377.

The following compounds were prepared in a manner similar to that described in Preparation 25:

25A: 1- [5-Bromo-6- (4-fluoro-benzyl) -3,3-dimethyl-2,3-dihydro-pyrrolo [3,2-b] pyridin-1-yl ]-Ethyl ketone, MS: [M + H] ⁺ = 377.

Preparation Example 26: 1- [6- (3-fluoro-benzyl) -3,3,5-trimethyl-2,3-dihydro-pyrrolo [3,2-b] pyridin-1-yl ]-Ethyl ketone (1- [6- (3-Fluoro-benzyl) -3,3,5-trimethyl-2,3-dihydro-pyrrolo [3,2-b] pyridin-1-yl] -ethanone)

In 1- [5-Bromo-6- (3-fluoro-benzyl) -3,3-dimethyl-2,3-dihydro-pyrrolo [3,2-b] pyridin-1-yl] -Ethyl ketone (which can be prepared as described in Preparation 25) (4.9 g, 13.0 mmol), LiBr (3.35 g, 39.0 mmol) and dichloride [1,3-bis (2,6- Diisopropylphenyl) imidazole-2-ylidene] (3-chloropyridine) palladium (II) ([1,3-bis (2,6-diisopropylphenyl) imidazol-2-ylidene) (3-chloropyridyl) palladium (II) dichloride (180 mg, 0.26 mmol) in degassed solution in THF (30 mL) and NMP (30 mL) with methylzinc chloride (in THF, 2M, 10 mL, 20 mmol), and the reaction mixture was stirred at room temperature overnight. The reaction mixture was poured into water (20 mL) and 5% aqueous citric acid solution (3 mL), and the product was extracted with toluene-EtOAc (1: 1, 2 x 40 mL). The organic phase was washed with brine, dried, filtered, and the solvent was evaporated to give the title compound (4.05 g). MS: [M + H] ⁺ = 313.

Preparation Example 27: 1- [6- (3-fluoro-benzyl) -3,3,5-trimethyl-4-oxy-2,3-dihydro-pyrrolo [3,2-b] Pyridine-1-yl] -ethanone (1- [6- (3-Fluoro-benzyl) -3,3,5-trimethyl-4-oxy-2,3-dihydro-pyrrolo [3,2-b] pyridin -1-yl] -ethanone)

In 1- [6- (3-fluoro-benzyl) -3,3,5-trimethyl-2,3-dihydro-pyrrolo [3,2-b] pyridin-1-yl] -ethyl To a solution of ketone (which can be prepared as described in Preparation Example 26) (4.05 g, 13.0 mmol) in DCM (50 mL) was added m-chloro-perbenzoic acid (77%, (4.4 g, 19.5 mmol), and the reaction mixture was stirred at room temperature for 2 hours. Add Na ₂ S ₂ O ₃ (10%, 50 mL) and stir for 30 minutes. The product was extracted with DCM (3x40 mL), the organic layers were mixed together, washed with 1M NaOH, dried, filtered, and the solvent was evaporated to give the title compound (4.22 g). MS: [M + H] ⁺ = 329.

Preparation Example 28: 1-Ethylacetic acid-6- (3-fluoro-benzyl) -3,3-dimethyl-2,3-dihydro-1H-pyrrolo [3,2-b] pyridine Acyl acid 1-acetyl-6- (3-fluoro-benzyl) -3,3-dimethyl-2,3-dihydro-1H-pyrrolo [3,2-b] pyridin-5-ylmethyl ester)

1- [6- (3-fluoro-benzyl) -3,3,5-trimethyl-4-oxy-2,3-dihydro-pyrrolo [3,2-b] pyridine-1 - yl] - ethanone (which may be prepared as described in preparation 27) (4.22 g, 12.86 mmol) in acetic anhydride (25 mL) was heated in the at 110 ^o C 2 hours. The reaction mixture was allowed to cool, poured onto ice, and stirred for 2 hours. The use of Na ₂ CO ₃ and mixtures, and using DCM (3x30 mL) and the product extracted. The organic phase was dried, filtered, and the solvent was evaporated. The crude product was purified on silica and eluted with petrol-EtOAc 0-50% to give the title compound (3.49 g). MS: [M + H] ⁺ = 371.

Preparation Example 29: [6- (3-fluoro-benzyl) -3,3-dimethyl-2,3-dihydro-1H-pyrrolo [3,2-b] pyridin-5-yl]- Methanol ([6- (3-Fluoro-benzyl) -3,3-dimethyl-2,3-dihydro-1H-pyrrolo [3,2-b] pyridin-5-yl] -methanol)

1-Ethylacetate-6- (3-fluoro-benzyl) -3,3-dimethyl-2,3-dihydro-1H-pyrrolo [3,2-b] pyridine-5- Methyl methyl ester (which can be prepared as described in Preparation 28) (3.49 g, 9.43 mmol) and NaOH (6.0 g, 150 mmol) in a solution of EtOH (60 mL) and water (60 mL) was heated And reflux overnight. EtOH was evaporated, the pH was adjusted to pH = 8 using 5 M HCl, and the product was extracted with DCM (3x30 mL). The organic phase was dried, filtered, and the solvent was evaporated. The crude product was purified on silica and eluted with petrol-EtOAc 0-100% to give the title compound (2.04 g). MS: [M + H] ⁺ = 287.

The following compounds were prepared in a sequence similar to that described in Preparations 25-29:

[6- (2-fluoro-benzyl) -3,3-dimethyl-2,3-dihydro-1H-pyrrolo [3,2-b] pyridin-5-yl] -methanol, [M + H] ⁺ = 287.

[6- (4-fluoro-benzyl) -3,3-dimethyl-2,3-dihydro-1H-pyrrolo [3,2-b] pyridin-5-yl] -methanol, [M + H] ⁺ = 287.

[6- (2,4-difluoro-benzyl) -3,3-dimethyl-2,3-dihydro-1H-pyrrolo [3,2-b] pyridin-5-yl] -methanol , ¹ H NMR (400 MHz, Me-d3-OD): 7.22-7.12 (1H, m), 7.00-6.82 (2H, m), 6.67-6.59 (1H, m), 4.72-4.61 (2H, m) , 4.04 (2H, s), 1.34 (6H, s).

Production Example 30: 1- [6- (4-fluoro-benzyl) -3,3-dimethyl-2,3-dihydro-pyrrolo [3,2-b] pyridin-1-yl]- Ethyl ketone (1- [6- (4-Fluoro-benzyl) -3,3-dimethyl-2,3-dihydro-pyrrolo [3,2-b] pyridin-1-yl] -ethanone)

6- (4-fluoro-benzyl) -3,3-dimethyl-2,3-dihydro-1H-pyrrolo [3,2-b] pyridine (which can be Acetyl chloride (3.6 mL, 51 mmol) was added to a MeCN (130 mL) solution prepared in Preparation Example 15 (10.1 g, 39 mmol). The mixture was stirred at room temperature overnight and then evaporated in vacuo. The residue was distributed between CH ₂ Cl ₂ and aqueous 1N NaOH was partitioned. The CH ₂ Cl ₂ layer was dried (MgSO ₄ ) and evaporated to give the title compound (12.3 g) as a crystalline solid. MS: m / z = 299 (M + H ⁺ ) ⁺ .

Preparation Example 31: 1- [6- (4-fluoro-benzyl) -3,3-dimethyl-4-oxy-2,3-dihydro-pyrrolo [3,2-b] pyridine- 1-yl] -ethyl ketone (1- [6- (4-Fluoro-benzyl) -3,3-dimethyl-4-oxy-2,3-dihydro-pyrrolo [3,2-b] pyridin-1-yl ] -ethanone)

1- [6- (4-fluoro-benzyl) -3,3-dimethyl-2,3-dihydro-pyrrolo [3,2-b] pyridin-1-yl] -ethanone ( It can be prepared as described in Preparation 30) (12.2 g, 41 mmol) and mCPBA (77%, 12 g, ~ 53 mmol) dissolved in CH ₂ Cl ₂ (150 mL), and stirred for 3 hours . A 20% aqueous Na ₂ S ₂ O ₃ solution was then added and the mixture was stirred for 25 minutes. Washed with an aqueous solution and then 2 x 1N NaOH to aqueous layer was extracted a number of CH ₂ Cl _2, and then CH ₂ Cl ₂ layers were combined. The organic layer was dried (MgSO _4), and evaporated in vacuo to give the title compound (12 g), as a yellow crystalline solid. MS: m / z = 315 (M + H ⁺ ) ⁺ .

Preparation Example 32: 1-Ethylacetic acid-6- (4-fluoro-benzyl) -3,3-dimethyl-2,3-dihydro-1H-pyrrolo [3,2-b] pyridine Acyl acid 1-acetyl-6- (4-fluoro-benzyl) -3,3-dimethyl-2,3-dihydro-1H-pyrrolo [3,2-b] pyridin-5-yl ester )

1- [6- (4-fluoro-benzyl) -3,3-dimethyl-4-oxy-2,3-dihydro-pyrrolo [3,2-b] pyridin-1-yl ] -Ethyl ketone (which can be prepared as described in Preparation 31) (11.55 g, 37 mmol) was heated in Ac ₂ O (70 mL) for 5 hours. The mixture was then cooled and poured into ice / water (500 g). The mixture was stirred for 1 hour, and the resulting precipitate was collected by filtration to give the title compound (12.1 g, 92%) as a gray solid. MS: m / z = 357 (M + H ⁺ ) ⁺ .

Production Example 33: 1-Ethyl-6- (4-fluoro-benzyl) -3,3-dimethyl-1,2,3,4-tetrahydro-pyrrolo [3,2-b] Pyridine-5-one (1-Acetyl-6- (4-fluoro-benzyl) -3,3-dimethyl-1,2,3,4-tetrahydro-pyrrolo [3,2-b] pyridin-5-one)

Make 1-ethylamyl-6- (4-fluoro-benzyl) -3,3-dimethyl-2,3-dihydro-1H-pyrrolo [3,2-b] pyridine-5- Ester (which can be prepared as described in Preparation 32) (6 g, 19 mmol) was suspended in EtOH (60 mL), and treated with 2N NaOH (42 mL) aqueous solution. The mixture was stirred overnight and then acidified with 5N aqueous HCl. The product was extracted with CH ₂ Cl ₂ and the organic layer was dried (MgSO ₄ ). Purification by SiO ₂ chromatography (eluted with 50-100% EtOAc / hexanes) gave a yellow solid. This was triturated with toluene and the solid was collected to give the title compound (2.4 g, 44%). MS: m / z = 315 (M + H ⁺ ) ⁺ .

The following compounds were prepared in a manner similar to that described in Preparations 30-33:

1-Ethyl-6- (2,4-difluoro-benzyl) -3,3-dimethyl-1,2,3,4-tetrahydro-pyrrolo [3,2-b] pyridine -5-one, MS: [M + H] ⁺ = 333.

1-Ethyl-6-butyl-3,3-dimethyl-1,2,3,4-tetrahydro-pyrrolo [3,2-b] pyridin-5-one, MS: [M + H] ⁺ = 263.

Preparation Example 34: 1-Ethyl-6- (4-fluoro-benzyl) -3,3,4-trimethyl-1,2,3,4-tetrahydro-pyrrolo [3,2- b] pyridin-5-one (1-Acetyl-6- (4-fluoro-benzyl) -3,3,4-trimethyl-1,2,3,4-tetrahydro-pyrrolo [3,2-b] pyridin- 5-one)

At about 0 ^o C in 1-acetyl-6- (4-fluoro - benzyl) -3,3-dimethyl-1,2,3,4-tetrahydro - pyrrolo [3,2 -b] pyridin-5-one (which can be prepared as described in Preparation 33) (3.1 g, 9.9 mmol) and K ₂ CO ₃ (2.7 g, 20 mmol) in DMF (30 mL) To the mixture was added methyl iodide (0.74 mL, 11.9 mmol). The mixture was stirred at room temperature for 5 hours, and then the mixture was partitioned between EtOAc and water. The EtOAc layer was washed with brine, and dried (MgSO _4). Purified by SiO ₂ chromatography (eluted with 0-10% MeOH / EtOAc) to give the title compound (960 mg, 29%) as a colorless crystalline solid. MS: [M + H] ⁺ = 329.

Preparation Example 35: 6- (4-fluoro-benzyl) -3,3,4-trimethyl-1,2,3,4-tetrahydro-pyrrolo [3,2-b] pyridine-5- Ketone (6- (4-Fluoro-benzyl) -3,3,4-trimethyl-1,2,3,4-tetrahydro-pyrrolo [3,2-b] pyridin-5-one)

1-Ethyl-6- (4-fluoro-benzyl) -3,3,4-trimethyl-1,2,3,4-tetrahydro-pyrrolo [3,2-b] pyridine -5-one (which can be prepared as described in Preparation 34) (960 mg, 2.9 mmol) is dissolved in a mixture of EtOH (10 mL) and 5N aqueous HCl (10 mL), and at 95 ^o C and Heat under N ₂ gas for 1 hour. The mixture was then allowed to cool and concentrated in vacuo. Ice and concentrated NH ₃ aqueous solution were added, and the resulting aqueous mixture was extracted with CH ₂ Cl ₂ . The CH ₂ Cl ₂ solution was dried (MgSO ₄ ) and evaporated to give the title compound, which was used immediately. MS: [M + H] ⁺ = 287.

The following compounds were prepared in a manner similar to that described in Preparations 30-35:

6- (2,4-difluoro-benzyl) -3,3,4-trimethyl-1,2,3,4-tetrahydro-pyrrolo [3,2-b] pyridin-5-one , MS: [M + H] ⁺ = 269.

The following compounds were prepared in a manner similar to that described in Preparation 35:

6- (4-fluoro-benzyl) -3,3, -dimethyl-1,2,3,4-tetrahydro-pyrrolo [3,2-b] pyridin-5-one, MS: [ M + H] ⁺ = 273.

6-butyl-3,3-dimethyl-1,2,3,4-tetrahydro-pyrrolo [3,2-b] pyridin-5-one, MS: [M + H] ⁺ = 221.

6-[(2,4-difluorophenyl) methyl] -3,3-dimethyl-5- pendantoxy-1H, 2H, 3H, 4H, 5H-pyrrolo [3,2-b] Pyridine, MS: [M + H] ⁺ = 291.

Preparation Example 36: 6- (2-fluoro-benzyl) -3,3,4-trimethyl-5- pendantoxy-2,3,4,5-tetrahydro-pyrrolo [3,2- b] Pyridine-1-carboxylic acid third butyl ester (6- (2-Fluoro-benzyl) -3,3,4-trimethyl-5-oxo-2,3,4,5-tetrahydro-pyrrolo [3,2 -b] pyridine-1-carboxylic acid tert-butyl ester)

Using a method similar to that described in Preparation 34 from 6-[(2-fluorophenyl) methyl] -3,3-dimethyl-5- pendantoxy-1H, 2H, 3H, 4H, 5H-pyrrole Preparation of benzo [3,2-b] pyridine-1-carboxylic acid tert-butyl ester. MS: [M + H] ⁺ = 387.

Preparation Example 37: 6- (2-fluoro-benzyl) -3,3,4-trimethyl-1,2,3,4-tetrahydro-pyrrolo [3,2-b] pyridine-5- Ketone (6- (2-Fluoro-benzyl) -3,3,4-trimethyl-1,2,3,4-tetrahydro-pyrrolo [3,2-b] pyridin-5-one)

Using a method similar to that described in Preparation 34 from 6- (2-fluoro-benzyl) -3,3,4-trimethyl-5- pendantoxy-2,3,4,5-tetrahydro-pyrrolo [3,2-b] Pyridine-1-carboxylic acid tert-butyl ester (which can be prepared as described in Preparation Example 36). MS: [M + H] ⁺ = 287.

Production Example 38: 1- [6- (4-fluoro-benzyl) -3,3-dimethyl-5-vinyl-2,3-dihydro-pyrrolo [3,2-b] pyridine- 1-yl] -ethyl ketone (1- [6- (4-Fluoro-benzyl) -3,3-dimethyl-5-vinyl-2,3-dihydro-pyrrolo [3,2-b] pyridin-1-yl ] -ethanone)

1- [5-Bromo-6- (4-fluoro-benzyl) -3,3-dimethyl-2,3-dihydro-pyrrolo [3,2-b] pyridin-1-yl] -Ethyl ketone (which can be prepared as described in Preparation Example 25A) (7.64 g, 20.27 mmol), tributylvinyltin (6.22 mL, 21.28 mmol) and bis (three third Butylphosphine) palladium (0) (bis (tri-tert-butylphosphine) palladium (0)) (0.104 g, 0.20 mmol) was dissolved in toluene (39 mL). After degassing with nitrogen, the reaction was heated to 85 ^o C for 2 hours. The reaction mixture was concentrated in vacuo, and the crude product was purified by column chromatography on silica gel (gradient elution, 0-100%, ethyl acetate / petrol 40-60 ^o C) to give the title compound ( 3.64 g). MS: [M + H] ⁺ = 325.

Preparation Example 39: (RS) -1- [5- (1,2-dihydroxy-ethyl) -6- (4-fluoro-benzyl) -3,3-dimethyl-2,3-di Hydrogen-pyrrolo [3,2-b] pyridin-1-yl] -ethanone ((RS) -1- [5- (1,2-Dihydroxy-ethyl) -6- (4-fluoro-benzyl)- 3,3-dimethyl-2,3-dihydro-pyrrolo [3,2-b] pyridin-1-yl] -ethanone)

1- [6- (4-fluoro-benzyl) -3,3-dimethyl-5-vinyl-2,3-dihydro-pyrrolo [3,2-b] pyridin-1-yl ]-Ethyl ketone (which can be prepared as described in Preparation Example 38) (3.64 g, 11.23 mmol) was placed in a mixture of acetone (76 mL) and water (8.5 mL) and an aqueous sodium hydroxide solution (2.5 M , 13.48 mL, 11.23 mmol), and the reaction was cooled to 0 ^o C (ice bath). Potassium permanganate (1.78 g, 11.23 moles) was added to the reaction, and stirring was continued for 1 hour. The reaction was allowed to warm to room temperature and stirred for 20 hours. Additional potassium permanganate (1.77 g, 33.7 mmol) was added, and the reaction was filtered through celite and washed with acetone and water. The filtrate was concentrated to give an aqueous mixture, which was extracted with ethyl acetate (3x). The organic layers were mixed together, dried over sodium sulfate, filtered, and concentrated in vacuo. The crude product was purified silica gel column chromatography (gradient elution, 0-100% ethyl acetate / petrol 40-60 ^o C), to give the title compound (1.5 g). MS: [M + H] ⁺ = 359.

Chiral purification

(RS) -1- [5- (1,2-Dihydroxy-ethyl) -6- (4-fluoro-benzyl) -3,3-dimethyl-2,3-dihydro-pyrrole [3,2-b] pyridin-1-yl] -ethanone (which can be prepared as described in Preparation 39) (1.5 g) was prepared by chiral preparative HPLC (ChiralPAK AD-H , Heptane / ethanol) to obtain 39A (R or S) -1- [5- (1,2-dihydroxy-ethyl) -6- (4-fluoro-benzyl) -3,3-di Methyl-2,3-dihydro-pyrrolo [3,2-b] pyridin-1-yl] -ethanone (fast-flowing isomer) (0.5 g) and 38B (R or S) -1- [5- (1,2-dihydroxy-ethyl) -6- (4-fluoro-benzyl) -3,3-dimethyl-2,3-dihydro-pyrrolo [3,2-b ] Pyridin-1-yl] -ethanone (slowly flowing isomer) (0.6 g).

Preparation Example 40: (RS) -1- [6- (4-fluoro-benzyl) -3,3-dimethyl-2,3-dihydro-1H-pyrrolo [3,2-b] pyridine -5-yl] -ethane-1,2-diol ((RS) -1- [6- (4-Fluoro-benzyl) -3,3-dimethyl-2,3-dihydro-1H-pyrrolo [3, 2-b] pyridin-5-yl] -ethane-1,2-diol)

(RS) -1- [5- (1,2-Dihydroxy-ethyl) -6- (4-fluoro-benzyl) -3,3-dimethyl-2,3-dihydro-pyrrole Benzo [3,2-b] pyridin-1-yl] -ethanone (which can be prepared as described in Preparation Example 39) (0.250 mg, 0.70 mmol) dissolved in ethanol (4.37 mL) and water (4.37 mL )in. Sodium hydroxide (0.447 g, 11.2 mmol) was added and the reaction was heated at reflux for 4 hours. After cooling to room temperature, the reaction mixture was concentrated. Water was added and the aqueous phase was extracted with ethyl acetate (3x). The organic extracts were mixed together, dried over sodium sulfate, filtered and concentrated to give the title compound (171 mg). MS: [M + H] ⁺ = 317.

The following compounds were prepared in a manner similar to that described in Preparation 40:

40A: (R or S) -1- [6- (4-fluoro-benzyl) -3,3-dimethyl-2,3-dihydro-1H-pyrrolo [3,2-b] pyridine -5-yl] -ethane-1,2-diol, from slower eluting isomer 39B. MS: [M + H] ⁺ = 317.

40B: (R or S) -1- [6- (4-fluoro-benzyl) -3,3-dimethyl-2,3-dihydro-1H-pyrrolo [3,2-b] pyridine -5-yl] -2-methoxy-ethanol, derived from the faster eluting precursor, MS: [M + H] ⁺ = 331.

40C: (R or S) -1- [6- (4-fluoro-benzyl) -3,3-dimethyl-2,3-dihydro-1H-pyrrolo [3,2-b] pyridine -5-yl] -2-methoxy-ethanol, precursor from slower elution, MS: [M + H] ⁺ = 331.

40D: (R or S) -2- [6- (4-fluoro-benzyl) -3,3-dimethyl-2,3-dihydro-1H-pyrrolo [3,2-b] pyridine -5-yl] -2-methoxy-ethanol, derived from the faster eluting precursor, MS: [M + H] ⁺ = 331.

40E: (R or S) -2- [6- (4-fluoro-benzyl) -3,3-dimethyl-2,3-dihydro-1H-pyrrolo [3,2-b] pyridine -5-yl] -2-methoxy-ethanol (from the precursor of the slower elution), MS: [M + H] ⁺ = 331.

Preparation Example 41: (RS) -Methanesulfonic acid 2- [1-ethylfluorenyl-6- (4-fluoro-benzyl) -3,3-dimethyl-2,3-dihydro-1H-pyrrole Benz [3,2-b] pyridin-5-yl] -2-hydroxy-ethyl ester ((RS) -Methanesulfonic acid 2- [1-acetyl-6- (4-fluoro-benzyl) -3,3-dimethyl -2,3-dihydro-1H-pyrrolo [3,2-b] pyridin-5-yl] -2-hydroxy-ethyl ester)

Was cooled to 0 ^o C in the (RS) -1- [5- (1,2- dihydroxy - ethyl) -6- (4-fluoro - benzyl) -3,3-dimethyl-2, 3-dihydro-pyrrolo [3,2-b] pyridin-1-yl] -ethanone (which can be prepared as described in Preparation 39) (1.48 g, 4.13 moles) in dichloromethane (20.7 mL ) To the mixture was added triethylamine (0.502 g, 4.96 mmol) and methanesulfonyl chloride (0.34 mL, 4.34 mmol). The reaction was warmed to room temperature and stirred for 2 hours. The reaction mixture was poured into water and extracted with DCM (3x). The organic extracts were mixed together, dried over sodium sulfate, filtered and concentrated in vacuo. Purified by silica gel column chromatography in the crude product (gradient elution, 0-100% ethyl acetate / petrol 40-60 ^o C), to give the title compound (1.25 g) MS: [M + H] + = 437.

Preparation Example 42: 1- [6- (4-fluoro-benzyl) -5- (1-hydroxy-2-methoxy-ethyl) -3,3-dimethyl-2,3-dihydro -Pyrrolo [3,2-b] pyridin-1-yl] -ethanone (1- [6- (4-Fluoro-benzyl) -5- (1-hydroxy-2-methoxy-ethyl) -3,3 -dimethyl-2,3-dihydro-pyrrolo [3,2-b] pyridin-1-yl] -ethanone) (42A) and 1- [6- (4-fluoro-benzyl) -5- (2- Hydroxy-1-methoxy-ethyl) -3,3-dimethyl-2,3-dihydro-pyrrolo [3,2-b] pyridin-1-yl] -ethanone (1- [6 -(4-fluoro-benzyl) -5- (2-hydroxy-1-methoxy-ethyl) -3,3-dimethyl-2,3-dihydro-pyrrolo [3,2-b] pyridin-1-yl]- ethanone) (42B)

In (RS) -methanesulfonic acid 2- [1-ethylfluorenyl-6- (4-fluoro-benzyl) -3,3-dimethyl-2,3-dihydro-1H-pyrrolo [3 , 2-b] pyridin-5-yl] -2-hydroxy-ethyl ester (which can be prepared as described in Preparation Example 41) (1.24 g, 2.84 mmol) was added to a mixture of methanol (9.48 mL) A mixture of sodium methoxide (25%) in methanol (1.23 mL, 5.69 mmol). After stirring for 6 hours, an additional mixture of sodium methoxide (25%) in methanol (1.23 mL) was added. The mixture was stirred for 18 hours, and then a mixture of sodium methoxide (25%) in methanol (1.23 mL) was added. Stir for another 22 hours, add water, and extract the reaction mixture with ethyl acetate (3x). The organic extracts were mixed together and concentrated in vacuo, and the crude product was purified by silica gel column chromatography (gradient elution, 0-100%, ethyl acetate / petrol 40-60 ^o C), and the two title compounds were individually obtained as external Racemic mixtures. Palm HPLC separation was performed as follows:

42A: ADH column, 80/20 heptane / ethanol, 0.2% DEA, 42A1 and 42A2 were obtained with faster elution.

42B: LUX-2 column, 80/20 heptane / ethanol, 0.2% DEA, to obtain faster eluted 42B1 [1H NMR (400 MHz, Me-d3-OD): 8.18 (1H, s), 7.19 ( 2H, dd), 7.03 (2H, t), 4.66 (1H, dd), 4.21-4.05 (2H, m), 4.05-3.82 (3H, m), 3.63 (1H, dd), 3.13 (3H, s) , 2.24 (3H, s), 1.42 (6H, s)] and the slower 42B2.

Preparation Example 43: (6-Methoxy-4-methyl-pyridin-3-yl) -carbamic acid tert- butyl ester)

In 5-amino-2-methoxy-4-picoline (5.0 g, 36.2 mmol) in THF (80 mL) and saturated Na ₂ CO ₃ To a solution of an aqueous solution (20 mL) was added di-tert-butyl-dicarbonate (7.9 g, 36.2 mmol), and the reaction mixture was stirred overnight. The reaction mixture was concentrated, extracted with DCM, washed with brine, dried, filtered, and the solvent was evaporated to give the title compound (8.8 g). MS: [M + H] ⁺ = 239.

Preparation Example 44: (5-tert-Butoxycarbonylamino-2-methoxy-pyridin-4-yl) -acetic acid )

(6-methoxy-4-methyl-pyridin-3-yl) -carbamic acid third butyl ester (which can be prepared as described in Preparation Example 43) at -78 ^o C (2.8 g, 11.9 Millimoles) were dissolved in THF (100 mL) and a second butyl lithium (1.4 M in cyclohexane, 28 mL, 39.3 millimoles) was added. The reaction mixture was stirred for 10 minutes, and then a bubble of CO _{2 was} bubbled through the cannula for 1 hour (1 h). Wait for the reaction mixture to warm to room temperature and quench with 2N HCl. The pH was adjusted to pH = 4 using 1 N NaOH, and the product was extracted with EtOAc. The organic layer was washed with brine, dried, filtered, and the solvent was evaporated to give the title compound (4.4 g). MS: [M + H] ⁺ = 283.

Preparation Example 45: 5-Methoxy-2-oxo-2,3-dihydro-pyrrolo [2,3-c] pyridine-1-carboxylic acid third butyl ester (5-Methoxy-2-oxo -2,3-dihydro-pyrrolo [2,3-c] pyridine-1-carboxylic acid tert-butyl ester)

(5-Third-butoxycarbonylamino-2-methoxy-pyridin-4-yl) -acetic acid (which can be prepared as described in Preparation 44) (3.4 g, 11.9 mmol) A mixture of propyl-ethyl-amine (4.6 mL, 26.18 mmol), EDC (2.5 g, 13.09 mmol) and HOAt (1.78 g, 13.09 mmol) in DCM (50 mL) was stirred for 3 hours . The reaction mixture with saturated aqueous NaHCO _3, water, brine, then dried, filtered, and the solvent was evaporated. The crude product was purified using silica chromatography, which was eluted with petrol-EtOAc 0-50% to give the title compound (2.2 g). MS: [M + H] ⁺ = 265.

Preparation Example 46: 5-methoxy-3,3-dimethyl-2- pendantoxy-2,3-dihydro-pyrrolo [2,3-c] pyridine-1-carboxylic acid third butyl ester (5-Methoxy-3,3-dimethyl-2-oxo-2,3-dihydro-pyrrolo [2,3-c] pyridine-1-carboxylic acid tert-butyl ester)

5-methoxy-2- pendantoxy-2,3-dihydro-pyrrolo [2,3-c] pyridine-1-carboxylic acid third butyl ester (which can be prepared as described in Preparation Example 45 ) (1.94 g, 7.35 mmol), K ₂ CO ₃ (2.33 g, 18.57 mmol) and methyl iodide (1.14 mL, 18.57 mmol) in acetone (25 mL) were heated at reflux for 3 hours. The reaction mixture was allowed to cool, the solvent was evaporated, and the residue was partitioned between water and DCM. The organic phase was dried, filtered, and the solvent was evaporated. The crude product was purified using silica chromatography, which was eluted with petrol-EtOAc 0-20% to give the title compound (1.47 g). MS: [M + H] ⁺ = 293.

Preparation Example 47: 6- (4-fluoro-benzyl) -3,3-dimethyl-2,5-dioxo-2,3,5,6-tetrahydro-pyrrolo [2,3 -c] pyridine-1-carboxylic acid tert-butyl ester (6- (4-Fluoro-benzyl) -3,3-dimethyl-2,5-dioxo-2,3,5,6-tetrahydro-pyrrolo [2, 3-c] pyridine-1-carboxylic acid tert-butyl ester)

5-methoxy-3,3-dimethyl-2- pendantoxy-2,3-dihydro-pyrrolo [2,3-c] pyridine-1-carboxylic acid third butyl ester (which may Prepared as described in Preparation 46) (1.43 g, 4.9 mmol), NaI (1.47 g, 9.8 mmol) and 4-fluorobenzyl bromide (0.67 mL, 5.4 mmol) Ear) The mixture in acetonitrile (50 mL) was heated at reflux for 5 hours, stirred at room temperature overnight, and heated at reflux for another 6 hours. The reaction mixture was cooled, poured into 10% Na ₂ S ₂ O ₃ aqueous solution, extracted with DCM, the organic phase was dried, filtered, and the solvent was evaporated. The crude product was purified using silica chromatography, which was eluted with petrol-EtOAc 0-100% to give the title compound (910 mg). MS: [M + H] ⁺ = 387.

The following compounds were prepared in a similar manner to that described in Preparation 47:

47A: 6- (2,4-difluoro-benzyl) -3,3-dimethyl-2,5-dioxo-2,3,5,6-tetrahydro-pyrrolo [2, 3-c] Pyridine-1-carboxylic acid tert-butyl ester. MS: [M + H] ⁺ = 405.

Preparation Example 48: 6- (4-fluoro-benzyl) -3,3-dimethyl-1,6-dihydro-3H-pyrrolo [2,3-c] pyridine-2,5-dione (6- (4-Fluoro-benzyl) -3,3-dimethyl-1,6-dihydro-3H-pyrrolo [2,3-c] pyridine-2,5-dione)

6- (4-Fluoro-benzyl) -3,3-dimethyl-2,5-dioxo-2,3,5,6-tetrahydro-pyrrolo [2,3-c] A solution of pyridine-1-carboxylic acid third butyl ester (which can be prepared as described in Preparation 47) (910 mg, 2.36 mmol) in TFA (5 mL) and DCM (5 mL) was stirred for 1 hour. The solvent was evaporated and the residue was partitioned between DCM and saturated _aqueous NaHC03, and the organic phase was dried, the solvent was evaporated to give the title compound (0.67 g). MS: [M + H] ⁺ = 287.

Preparation Example 49: 6- (4-fluoro-benzyl) -3,3-dimethyl-1,2,3,6-tetrahydro-pyrrolo [2,3-c] pyridin-5-one ( 6- (4-fluoro-benzyl) -3,3-dimethyl-1,2,3,6-tetrahydro-pyrrolo [2,3-c] pyridin-5-one)

In 6- (4-fluoro-benzyl) -3,3-dimethyl-1,6-dihydro-3H-pyrrolo [2,3-c] pyridine-2,5-dione (which can Prepared as described in Preparation 48) (526 mg, 1.84 mmol) in a solution of THF (30 mL), add a solution of BH ₃ .Me ₂ S (2M, 9.7 mL, 18.4 mmol), and The mixture was heated at reflux for 3 hours. Cool, carefully add MeOH (10 mL), and heat to reflux for 2 hours. The solvent was evaporated and the residue was partitioned between DCM and saturated aqueous NaHCO _3. The organic phase was dried, filtered, and the solvent was evaporated to give the title compound (494 mg). It was used without purification. MS: [M + H] ⁺ = 273.

The following compounds were prepared in a procedure similar to that described in Preparation 49:

49A: 6- (2,4-difluoro-benzyl) -3,3,4-trimethyl-1,2,3,6-tetrahydro-pyrrolo [2,3-c] pyridine-5 -Ketone, MS: [M + H] ⁺ = 305.

The following compounds were prepared by procedures similar to those described in Preparations 47-49:

49B: 6- (2,4-difluoro-benzyl) -3,3-dimethyl-1,2,3,6-tetrahydro-pyrrolo [2,3-c] pyridin-5-one , MS: [M + H] ⁺ = 291.

Preparation Example 50: 4-Amino-1- (2-chloro-ethenyl) -6- (2,4-difluoro-benzyl) -3,3-dimethyl-1,2,3, 4-tetrahydro-pyrrolo [3,2-b] pyridin-5-one (4-Amino-1- (2-chloro-acetyl) -6- (2,4-difluoro-benzyl) -3,3- dimethyl-1,2,3,4-tetrahydro-pyrrolo [3,2-b] pyridin-5-one)

In 1- (2-chloro-ethylamyl) -6- (2,4-difluoro-benzyl) -3,3-dimethyl-1,2,3,4-tetrahydro-pyrrolo [ 3,2-b] pyridin-5-one (which can be prepared as described in Preparation 18D) (117 mg, 0.32 mmol) was dissolved in DMF (2 mL) and potassium carbonate (88 mg, 0.64 Millimolar) and o- (2,4-dinitrophenyl) -hydroxylamine (95 mg, 0.48 millimolar). The resulting mixture was stirred at room temperature for 2 hours. 1 M aqueous sodium hydroxide (5 mL) was added, and the mixture was extracted with EtOAc (2 x 10 mL). Parts of the organic washed with water (organic fractions), dried MgSO _4, and concentrated. The residue was purified using column chromatography using 20-65% EtOAc in petrol as the eluent to give the title compound (79 mg) as an orange solid. MS: [M + H] ⁺ = 382.

The following compounds were prepared in a procedure similar to that described in Preparation 50:

50A: 4-Amino-1- (2-chloro-ethenyl) -6- (4-fluoro-benzyl) -3,3-dimethyl-1,2,3,4-tetrahydro- Pyrrolop [3,2-b] pyridin-5-one, MS: [M + H] ⁺ = 364.

50B: 4-amino-1- (2-chloro-ethylfluorenyl) -6-butyl-3,3-dimethyl-1,2,3,4-tetrahydro-pyrrolo [3,2- b] Pyridin-5-one, MS: [M + H] ⁺ = 312.

Preparation Example 51: 4-bromo-6- (2,4-difluoro-benzyl) -3,3-dimethyl-2,5-dioxo-2,3,5,6-tetrahydro -Pyrrolo [2,3-c] pyridine-1-carboxylic acid tert-butyl ester (4-Bromo-6- (2,4-difluoro-benzyl) -3,3-dimethyl-2,5-dioxo-2 , 3,5,6-tetrahydro-pyrrolo [2,3-c] pyridine-1-carboxylic acid tert-butyl ester)

Add N-bromosuccinimide (529 mg, 2.97 mmol) to 6- (2,4-difluoro-benzyl) -3,3-dimethyl-2,5-dioxo- 2,3,5,6-tetrahydro-pyrrolo [2,3-c] pyridine-1-carboxylic acid tert-butyl ester (which can be prepared as described in Preparation Example 47A) (1.0 g, 2.47 mmol) ) Dissolved in DMF solution. The solution was stirred at 60 ° C for 1.5 hours. The reaction mixture was allowed to cool to room temperature, water was added, and the product was extracted with DCM (3x). The organic phase was collected, dried over Na ₂ SO _4, filtered, and concentrated in vacuo. The residue was purified by flash chromatography to obtain 1.1 g of the title compound as a yellow solid. MS: [M + H] ⁺ = 484.

Preparation Example 52: 6- (2,4-difluoro-benzyl) -3,3,4-trimethyl-1,6-dihydro-3H-pyrrolo [2,3-c] pyridine-2 , 5-dione (6- (2,4-Difluoro-benzyl) -3,3,4-trimethyl-1,6-dihydro-3H-pyrrolo [2,3-c] pyridine-2,5-dione)

Slowly add Me ₂ Zn in heptane (1M, 5.8 mL, 5.8 mmol) to 4-bromo-6- (2,4-difluoro-benzyl) -3,3-dimethyl-2 3,5-Dioxo-2,3,5,6-tetrahydro-pyrrolo [2,3-c] pyridine-1-carboxylic acid third butyl ester (which can be prepared as described in Preparation Example 51) (935 mg, 1.93 mmol) and Pd (dppf) ₂ Cl ₂ (282 mg, 0.38 mmol) were dissolved in a solution of dioxane (10 mL). The reaction mixture was stirred in a sealed vessel at 70 ° C under N ₂ for 1 hour. A second aliquot of Me ₂ Zn (5.8 mL, 5.8 mmol) was then added and stirring was continued for 2 hours. The reaction mixture was cooled to room temperature, saturated aqueous NaHCO ₃ to quench it, and extracted with DCM. The organic phase was dried over Na ₂ SO _4, filtered, and concentrated in vacuo. The crude product was purified by flash chromatography to give 200 mg of the title compound as a yellow semi-solid. MS: [M + H] ⁺ = 319.

Preparation Example 53: 5-Methoxy-3,3-dimethyl-1,3-dihydro-pyrrolo [2,3-c] pyridin-2-one (5-Methoxy-3,3-dimethyl- 1,3-dihydro-pyrrolo [2,3-c] pyridin-2-one)

Using a procedure similar to that described in Preparation 48, from 5-methoxy-3,3-dimethyl-2- pendantoxy-2,3-dihydro-pyrrolo [2,3-c] pyridine- Preparation of 1-carboxylic acid third butyl ester. MS: [M + H] ⁺ = 193.

Preparation Example 54: 5-Methoxy-3,3-dimethyl-2,3-dihydro-1H-pyrrolo [2,3-c] pyridine (5-Methoxy-3,3-dimethyl-2, 3-dihydro-1H-pyrrolo [2,3-c] pyridine)

In 5-methoxy-3,3-dimethyl-1,3-dihydro-pyrrolo [2,3-c] pyridin-2-one (which can be prepared as described in Preparation 53) (2.8 g, 14.6 mmol) was dissolved in THF (60 mL) was added a solution of BH ₃ .THF solution (1M, 150 mL, 150 mmol), and the mixture was stirred overnight at room temperature. Carefully add MeOH (50 mL) and heat to reflux for 1 hour. The solvent was evaporated and the residue was partitioned between DCM and saturated aqueous NaHCO _3. The organic phase was dried, filtered, and the solvent was evaporated. The crude product was purified using silica chromatography, which was eluted with petrol-EtOAc 0-60% to give the title compound (2.27 g). MS: [M + H] ⁺ = 179.

Preparation Example 55: 5-Methoxy-3,3-dimethyl-2,3-dihydro-pyrrolo [2,3-c] pyridine-1-carboxylic acid third butyl ester (5-Methoxy-3 , 3-dimethyl-2,3-dihydro-pyrrolo [2,3-c] pyridine-1-carboxylic acid tert-butyl ester)

In 5-methoxy-3,3-dimethyl-2,3-dihydro-1H-pyrrolo [2,3-c] pyridine (which can be prepared as described in Preparation 54) (534 mg, 3.0 millimolar) in a solution of THF (10 mL) and saturated Na ₂ CO ₃ aqueous solution (4 mL) was added di-tert-butyl dicarbonate (780 mg, 3.6 millimolar), and the reaction mixture was stirred Overnight, then diluted with water, extracted with EtOAc, washed with brine, dried, and evaporated the solvent to give the title compound (760 mg). MS: [M + H] ⁺ = 279.

Preparation Example 56: 6-((E) -but-2-enyl) -3,3-dimethyl-5- pendantoxy-2,3,5,6-tetrahydro-pyrrolo [2,3 -c] Pyridine-1-carboxylic acid tert-butyl ester (6-((E) -But-2-enyl) -3,3-dimethyl-5-oxo-2,3,5,6-tetrahydro-pyrrolo [ 2,3-c] pyridine-1-carboxylic acid tert-butyl ester)

5-methoxy-3,3-dimethyl-2,3-dihydro-pyrrolo [2,3-c] pyridine-1-carboxylic acid third butyl ester (which can be as described in Preparation Example 55 And prepared) (760 mg, 2.7 millimoles), NaI (410 mg, 2.7 millimoles), and a mixture of crotyl bromide (0.33 mL, 3.24 millimoles) in acetonitrile (25 mL) Heat to reflux for 5 hours. The reaction mixture was allowed to cool, poured onto 10% Na ₂ S ₂ O ₃ (on 10% Na ₂ S ₂ O ₃ ), extracted with DCM, the organic phase was dried, filtered, and the solvent was evaporated. The crude product was purified using silica chromatography, which was eluted with petrol-EtOAc 0-70% to give the title compound (433 mg). MS: [M + H] ⁺ = 319.

Preparation Example 57: 6-butyl-3,3-dimethyl-5- pendantoxy-2,3,5,6-tetrahydro-pyrrolo [2,3-c] pyridine-1-carboxylic acid Tributyl ester (6-Butyl-3,3-dimethyl-5-oxo-2,3,5,6-tetrahydro-pyrrolo [2,3-c] pyridine-1-carboxylic acid tert-butyl ester)

6-((E) -but-2-enyl) -3,3-dimethyl-5- pendantoxy-2,3,5,6-tetrahydro-pyrrolo [2,3-c] Tertiary butyl pyridine-1-carboxylic acid (which can be prepared as described in Preparation 56) (433 mg, 1.36 mmol) and Pd / C (10%, 100 mg) in EtOH (15 mL) The mixture was hydrogenated for 1 hour. The catalyst was filtered, the filtrate was evaporated, and the residue was purified using silica gel chromatography, which was eluted with petrol-EtOAc 0-50% to give the title compound (387 mg). MS: [M + H] ⁺ = 321.

Preparation Example 58: 6-Butyl-3,3-dimethyl-1,2,3,6-tetrahydro-pyrrolo [2,3-c] pyridin-5-one (6-Butyl-3,3 -dimethyl-1,2,3,6-tetrahydro-pyrrolo [2,3-c] pyridin-5-one)

6-Butyl-3,3-dimethyl-5- pendantoxy-2,3,5,6-tetrahydro-pyrrolo [2,3-c] pyridine-1-carboxylic acid tert-butyl ester (380 mg, 1.19 mmol) dissolved in TFA (5 mL) and DCM (5 mL) and stirred for 1 hour. The solvent was evaporated, the residue was partitioned between DCM and saturated aqueous NaHCO ₃ solution, the organic phase was dried, the solvent was evaporated, and the residue was purified using silica gel chromatography, which was eluted with petrol-EtOAc 0-100% To give the title compound (170 mg). MS: [M + H] ⁺ = 221.

Examples 1 to 37

The following steps illustrate the preparation of Examples 1 to 37 listed in the table below.

(2R, 5S) -4- (2- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrole Benzo [3,2-b] pyridin-1-yl} -2-oxoethyl) -2-methyl-5-{[((3R) -3-methylmorpholin-4-yl] methyl } Piperazine-1-carboxylic acid tert-butyl ester (tert-butyl (2R, 5S) -4- (2- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl -1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-oxoethyl) -2-methyl-5-{[(3R) -3-methylmorpholin-4-yl] methyl} piperazine -1-carboxylate) (0.47 g) , ethyl acetate (10 mL), and HCl - dioxane (dioxane) (4 M; mixture 10 mL) was stirred for 18 hours at the 20 ^o C, the resulting solid was collected by filtration To give 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] Pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} piperazin-1-yl] Ethan-1-one dihydrochloride (Example 2, 0.43 g).

The compounds listed in the table below were prepared by similar and / or methods similar to those described above from the corresponding N-Boc protected derivatives, and any significant changes are described below. The precursors of N-Boc protected derivatives are indicated (by preparation number or name) in the table below. The title compound is isolated directly as a free base or a suitable salt without further purification, or is purified, for example, using mass-oriented preparative HPLC, crystallization, or trituration.

1H NMR is generated at 400 MHz and is in Me-d3-OD, unless stated otherwise.

Example 38: 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b ] Pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} piperazin-1-yl ] Ethyl-1-one (1- {6-[(4-Fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1 -yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} piperazin-1-yl] ethan-1-one) (free base (Free Base))

1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridine -1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} piperazin-1-yl] ethyl 1-one dihydrochloride (1.00 g, 1.0 equivalent (eq.), 1.00 wt.) (Which can be prepared as described in Example 2) was fed to an RB flask and dissolved in water (10.0 mL, 10.0 vol , 10.00 wt.), And stirred under nitrogen at 18 to 23 ° C to obtain a straw coloured solution (pH = 4.73, T = 19.3 ° C). Ethyl acetate (10.0 mL, 10.0 vol) was added to the aqueous solution, and the biphasic mixture was stirred at 18 to 23 ° C for 5 minutes. The layers were separated and the aqueous layer (pH = 4.58, 19.6 ° C) was poured back into the flask. (Cautious) Effervescence was observed with the addition of sodium bicarbonate (388.2 mg, 3 x 1.05 eq., 0.4 wt.). The mixture was stirred for 20 minutes (pH = 7.51, 18.2 ° C), dichloromethane (5.0 mL, 5.0 vol) was added, and the mixture was stirred for another 5 minutes under the same conditions. The layers were separated, the dichloromethane layer was retained, and the aqueous layer (pH = 7.66, T = 17.7 ° C) was poured back into the flask. Dichloromethane (2 x 5.0 mL, 2 x 5.0 vol) was used for secondary extraction (pH = 8.25, T = 18.5 ° C & pH = 8.47, T = 18.3 ° C). The organic layers were mixed together with sodium sulfate (1.0 g, 1.0 wt.) Was dried, filtered, and concentrated to dryness under reduced pressure at 40 ° C (400 mbar). The concentrate was then dried at 40 ° C (<20 mbar) for 2 hours to obtain a white foam (850.2mg, 102% th., 100% corr. Input and output w / w assay), 94.3% w / w osfb (for TCNB), which contains ethyl acetate (3.4% w / w) and dichloromethane (0.8% w / w).

Example 39: 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b ] Pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} piperazin-1-yl ] Eth-1-one L-(+)-lactate (Form A)

1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridine -1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} piperazin-1-yl] ethyl A free base of 1-one (500.0 mg, 1.0 wt) (which can be prepared as described in Example 38) was fed into a 25 mL container and dissolved in ethanol (1.0 mL, 2.0 vol). L-(+)-lactic acid (106.2 mg, 1.0 eq) was added, and the contents of the container were stirred at 18 to 23 ° C. for 1 hour to obtain a yellow solution. After this time, TBME (9.0 mL, 18.0 vol) was fed into the container, and the mixture was continuously stirred at 18 to 23 ° C. The progress of salt crystallization was monitored by XRPD. After stirring at 18 to 23 ° C for 16 hours, lactate remained in the solution. After this time, the solution was concentrated to about ¼ of the original volume, and TBME (9.0 mL, 18.0 vol) was added to obtain a gummy solid and a clear supernatant, which were subjected to sonic vibration treatment and further stirred (in After about 20 hours at 18 to 23 ° C), it became a suspension of fine particles. Filtration was performed to isolate the solid and dried under a stream of nitrogen to give 365 mg of a white solid, identified as Form B by XRPD. The solid was dried in an oven at 40 to 45 ° C for 67 hours to obtain a white solid (325mg, 56% th.), 98.8% w / w oasfb (for TCNB), which contained TBME (1.0% w / w) and water (0.6% w / w) as determined by XRPD as Form A. The detailed characteristic data ( ¹ H NMR, XRPD, and DSC) of Example 39 are shown in FIGS. 1 to 3.

Example 40: 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b ] Pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} piperazin-1-yl ] Eth-1-one L-(+)-lactate (Form B)

The reaction was performed according to the procedure described in Example 39, but without drying in an oven to obtain a white solid (529.1mg, 90% th.), 96.1% w / w oasfb (for TCNB), which contained TBME (5.9% w / w) and Water (3.8% w / w) and Form X as determined by XRPD. Another procedure can be used for Example 40 to avoid the use of ethanol. The detailed characteristic data ( ¹ H NMR, XRPD and DSC) of Example 40 are shown in FIGS. 4 to 6.

Example 41: 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b ] Pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} piperazin-1-yl ] Eth-1-one sulfate (Form F)

1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridine -1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} piperazin-1-yl] ethyl A free base of 1-one (500.0 mg, 1.0 wt) (which can be prepared as described in Example 38) was charged into a 10 mL container and dissolved in ethanol (1.0 mL, 2.0 vol). A solution of sulfuric acid (103.2 mg, 1.0 eq.) In ethanol (4.0 mL, 8.0 vol) was added at 18 to 23 ° C. with stirring for 10 minutes to obtain a clear gel. The contents of the container were stirred at the same temperature for 1 hour, during which the gums were dissolved to give a yellow solution. Stirring was continued for 16 hours, resulting in a white suspension. The progress of salt crystallization was monitored by XRPD. Ethanol (2.0 mL, 4.0 vol) was then added to allow the suspension to properly mobilise, and the product was isolated by filtration and dried under a stream of nitrogen to give a white solid (465.2 mg, 79% th.), 94.9% w / w oasfb (for TCNB), which contains ethanol (2.9% w / w) and water (3.6% w / w), shows Form F as determined by XRPD. The detailed characteristic data ( ¹ H NMR, XRPD and DSC) of Example 41 are shown in FIGS. 7 to 9.

Example 42: 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b ] Pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} piperazin-1-yl ] Eth-1-one mesylate (Form B)

1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridine -1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} piperazin-1-yl] ethyl A free base of 1-one (500.0 mg, 1.0 wt) (which can be prepared as described in Example 38) was charged into a 25 mL container and dissolved in 2-propanol (2.5 mL, 5.0 vol). Methanesulfonic acid (276.0 mg, 3.0 eq.) Was added (a small amount of exotherm was observed), and the resulting oily-gummy mixture was stirred at 18 to 23 ° C. N-heptane (10.0 mL, 20.0 vol) was slowly added over a period of 10 minutes to obtain a white suspension and a small amount of gummy solid. The progress of salt crystallization was monitored by XRPD. Under mild conditions (stirring at 18 to 23 ° C for 3 days) the salt did not crystallize, so the temperature was increased to 40 to 45 ° C to obtain sticky gummy solids and clear supernatant ). This mixture was cooled to 18 to 23 ° C, mobilized with a spatula, and sonicated for 20 minutes to obtain a white suspension containing some gummy solids. The suspension was stirred at the same temperature for 20 hours, filtered, and dried under a stream of nitrogen to obtain a beige solid (402.9 mg, 63% th.), 99.0% w / w oasfb (for TCNB), which contained 2- Propanol (2.3% w / w), n-heptane (0.2% w / w), and water (1.9% w / w) showed Form B as determined by XRPD. The detailed characteristic data ( ¹ H NMR, XRPD and DSC) of Example 42 are shown in FIGS. 10 to 12.

Example 43: 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b ] Pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} piperazin-1-yl ] Eth-1-one L-(+)-lactate (Form C)

The first batch

1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridine- 1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} piperazin-1-yl] ethyl- The free base of 1-ketone (10.0 g, corr.) (Which can be prepared as described in Example 38) was dissolved in isopropyl acetate (80.0 mL, 8.0 vol) to give a pale yellow solution. A solid, anhydrous L-(+)-lactic acid (1.67 g, 1.0 eq.) Was charged into the same flask in one portion, and a small amount of gum was clearly visible at the bottom of the flask. The mixture was then quickly stirred to mobilise the colloid, the mixture spontaneously nucleated, and solids precipitated. A sample of the solid precipitate was analyzed at XRPD, which was consistent with Form B. N-heptane (12.0 vol) was added and the mixture was stirred at 40 ° C for 4 days under nitrogen, showing that the mixture would transform to Form C, and its progress was monitored by XRPD (Figure 13). The temperature of the mixture was increased to 55 ° C, and stirring was continued for 24 hours to complete the conversion (Figure 14). The product was isolated by filtration (quickly <0.5 minutes), washed with isopropyl acetate / n-butane (2.0 / 3.0, v / v, 5.0vol), and dried under a nitrogen stream for 20 hours to give the title compound as a white powder (8.89 g, 79% th.), 91.6% w / w (osfb), Form C, mp 172 ° C.

Second batch

1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridine -1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} piperazin-1-yl] ethyl The free base of 1-one (10.0 g, corr.) (Which can be prepared as described in Example 38) was dissolved in isopropyl acetate (60.0 mL, 6.0 vol) to give a pale yellow solution. A solution of anhydrous L-(+)-lactic acid (1.67 g, 1.0 eq.) In isopropyl acetate (10.0 mL, 1.0 vol) was added thereto. A line rinse of isopropyl acetate (10.0 mL, 1.0 vol) was performed, and the mixture was stirred at 18 to 23 ° C to obtain a pale yellow solution.

N-heptane (120 mL, 12.0 vol) was added dropwise over a period of 40 minutes, and a gum body was formed on the bottom of the flask. After stirring for 1 hour and 40 minutes, the appearance of the mixture improved, but gum gums still appeared on the bottom of the flask.

The mixture was stirred at 18 to 23 ° C for 16 hours, during which time the colloidal body was mobilised, forming a granular fine suspension. The suspension was filtered (rapid filtration) under nitrogen, and the filter cake was sampled and analyzed by XRPD (form C). The filter cake was washed with isopropyl acetate / n-heptane (2.0 / 3.0, v / v, 5.0 vol), sampled, analyzed by XRPD (Form C), and the cake was left on the filter under a nitrogen stream to pull dry. ) 16 hours. The product consists of a slightly off-white powdery solid (11.36 g, 91% corr.), Form C, 94.3% w / w (oasfb), Form C, mp 172 ° C and contains isopropyl acetate Ester (1.0% ww).

The detailed characteristic data ( ¹ H NMR, XRPD and DSC) of Example 43 are shown in FIGS. 15 to 17.

Example 44: 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b ] Pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} piperazin-1-yl ] Eth-1-one L-(+)-lactate (Form C)

step 1

(2R, 5S) -4- (2- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, cooled to <10 ° C , 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-oxoethyl) -2-methyl-5-{[((3R) -3-methylmorpholine- 4-Methyl} piperazine-1-carboxylic acid tert-butyl ester (1.0 wt) (which can be prepared as described in Preparation Example 19) in methanol (10 vol), slowly add 4M HCl-1, 4-Dioxane (1,4-dioxane) (3 vol) solution, followed by a line rinse of methanol (0.5 vol). The mixture was warmed to 15 to 25 ° C and stirred at this temperature for at least 12 hours. The reaction mixture was then warmed to 30 to 40 ° C and stirred until the reaction was complete by HPLC (generally> 2 hours). When the reaction is complete, the reaction solution is concentrated to dryness at up to 40 ° C. The residue was dissolved in purified water (8 vol) and washed with ethyl acetate (2 x 4 vol). The pH of the aqueous phase was adjusted to pH 12 to 13 using 4M NaOH (as required), followed by extraction with ethyl acetate (3 x 5 vol). The organic phases were mixed together and washed with sodium chloride solution (5 vol) and magnesium sulfate (1.0 wt) for at least 10 minutes. The solids were removed by filtration, and the filter cake was washed with ethyl acetate (2 x 2 vol). The filtrate was concentrated on a rotary evaporator at up to 40 ° C, the resulting concentrate was dissolved in methyl acetate (5 vol), and the solution was concentrated as described above to produce 1- {6-[(4-fluorobenzene (Methyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[(3R) -3-methylmorpholin-4-yl] methyl} piperazin-1-yl] ethan-1-one is a free base.

Step 2

1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] pyridine -1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} piperazin-1-yl] ethyl To a free-base methyl acetate (3 vol) solution of 1-one (1.0 wt) (which can be prepared as described in step 1), add methyl acetate (L-(+)-lactic acid (0.085 wt)) ( 0.75 vol) solution. Then add 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H, 2H, 3H-pyrrolo [3,2-b] Pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholin-4-yl] methyl} piperazin-1-yl] Seed slurry of aceto-1-one L-(+)-lactate (Form C) (0.01 wt) in methyl acetate (0.08 vol), then L-(+)-lactic acid ( 0.085 wt) methyl acetate (0.75 vol) solution, and a line rinse of methyl acetate (0.5 vol) was performed. The suspension was stirred for about 30 minutes, and then n-heptane (12.0 vol) was added over a period of at least 1 hour, and the temperature was maintained at 15 to 25 ° C. Maintain the mixture at 15 to 25 ° C and stir for at least 2 hours. The solids were removed by filtration, and the filter cake was washed with 2: 3 methyl acetate / n-butane (5 vol). Dry the material on the filter until it is suitable for processing, and then dry in an oven up to 80 ° C until the methyl acetate content is ≤ 0.5% w / w to obtain the title compound, off-white to Beige solid.

Example 45: Examples of Pharmaceutical Formulations

(i) Lozenge formula

A suitable amount of a compound of formula (I) (for example, 50-250 mg) is mixed with an appropriate diluent, disintegrant, compression agent, and / or glidant to prepare a formula containing the formula (I) A lozenge composition of a compound. One possibility is that the lozenge contains 50 mg of the compound, and 197 mg of lactose (BP) as a diluent, and 3 mg of magnesium stearate as a lubricant, in a known manner. Compress to form an ingot. The pressed tablets can be coated with a film.

(ii) Capsule formula

100-250 mg of the compound of formula (I) is mixed with an equivalent amount of lactose, and the resulting mixture is filled into standard hard gelatin capsules to prepare a capsule formula. Appropriate disintegrating adjuvants and / or glidants may be included as required.

(iii) Injection Formula I

A parenteral composition for injection can be prepared by dissolving a compound of formula (I) (for example, in the form of a salt) in water containing 10% propylene glycol to obtain an active compound concentration of 1.5% by weight. The solution is then made isotonic, filtered or terminally sterilized, filled into ampoules or vials, or pre-filled syringes, and sealed.

(iv) Injection Formula II

A parenteral composition for injection can be prepared by dissolving a compound of formula (I) (for example, in the form of a salt) (2 mg / ml) and mannitol (50 mg / ml) in water, and filtering or terminal sterilizing the solution Fill in a sealable 1 ml vial or ampoule, or a pre-filled syringe.

(v) Injection Formula III

A compound of formula (I) (e.g. in the form of a salt) is dissolved in water (2 mg / ml), and then isotonicity is adjusted to prepare an i.v. delivery formulation via injection or infusion. The vial is then sealed and autoclaved or filled into an ampoule or vial, or pre-filled into a syringe, filtered and sterilized and sealed.

(vi) Injection Formula IV

A compound of formula (I) (e.g. in the form of a salt) is dissolved in water containing a buffer (e.g. 0.2M acetate, pH 4.6) in an amount of 2 mg / ml (2 mg / ml) to prepare via injection or Infused intravenous delivery formula. Vials, ampoules, or pre-filled syringes are then sealed and autoclaving, or filter sterilized and sealed.

(vii) Subcutaneous or Intramuscular injection formula

The compound of formula (I) is mixed with pharmaceutical-grade corn oil to prepare a subcutaneous administration composition to obtain a concentration of 5-50 mg / ml. The composition is sterilized and filled into a suitable container.

(viii) Freeze-dried formula I

The formulated compound of formula (I) was dispensed into 50 ml vials and freeze-dried. During freeze-drying, the composition is frozen at -45 ºC using a one-step freezing protocol. Raise the temperature to –10 ºC for annealing, then lower the temperature to freeze at –45 ºC, then dry initially at +25 ºC for about 3400 minutes, and then gradually increase to 50 ºC for the second time Secondary drying. The pressure during the initial drying and the second drying was set at 80 millitor.

(ix) Freeze-dried Formulation II

The formulated compound of formula (I) or a salt thereof (as defined herein) is dispensed into a 50 ml vial and freeze-dried. During freeze-drying, the composition is frozen at -45 ºC using a one-step freezing protocol. Raise the temperature to –10 ºC for annealing, then lower the temperature to freeze at –45 ºC, then dry initially at +25 ºC for about 3400 minutes, and then gradually increase to 50 ºC for the second time Secondary drying. The pressure during the initial drying and the second drying was set at 80 millitor.

(x) Freeze-dried formula III for intravenous administration

The compound of formula (I) is dissolved in a buffer solution to prepare an aqueous buffered solution. The buffer solution is filtered to remove particulate matter and filled into a container (such as a glass vial of Type 1) and then partially sealed (for example, by a Fluorotec stopper). If the compound and formula are stable enough, the formula can be autoclaved at 121ºC for a suitable period of time. If the formulation is unstable to autoclave sterilization, it can be sterilized with a suitable filter and filled into a sterilized vial under sterilization. The solution was freeze-dried using a suitable cycle. When the freeze-drying cycle is complete, backfill the vial with nitrogen to atmospheric pressure, stopper the stopper, and securely close it (eg, using aluminum crimp). For intravenous administration, the lyophilized solid can be restored to its original state in a pharmaceutically acceptable diluent, such as 0.9% saline or 5% dextrose. This solution can be administered in its original form, or prior to administration, into an infusion bag (containing a pharmaceutically acceptable diluent, such as 0.9% saline or 5% dextrose).

(xi) Active pharmaceutical ingredients in bottles

A bottle or vial is filled with a compound of formula (I) to prepare a composition for oral administration. The composition is then reduced with a suitable diluent such as water, fruit juice or a commercially available excipient such as OraSweet or Syrspend. The reducing solution can be taken into a measuring cup or an oral syringe for administration.

Biological Assays

XIAP, cIAP-1 and cIAP-2 BIR3 regions

Fusion of human XIAP (residues 252-350) with His-tag, fusion of human cIAP-1 (residues 267-363) with GST-tag, and fusion of human cIAP-2 (residues 244-337) with His-tag The recombinant BIR3 region was overexpressed in E. coli cells grown in TB medium. Proteins were isolated from the cell lysate using Ni-NTA affinity chromatography (XIAP / cIAP-2) or glutathione sepharase 4B affinity chromatography (cIAP-1). The affinity tags of XIAP and cIAP-1 were cut with thrombin in 25mM HEPES pH 7.5, 100mM NaCl, 50uM Zn (OAc) ₂ and 1mM Ca (OAc) ₂ and then analyzed by particle size sieve BIR3 region was purified by size-exclusion chromatography. His-tag of cIAP-2 has not been excised and due to aggregation induced covalent self-oligomerization issues, the protein is not concentrated above 3 mg / ml. The purified protein was stored in 25 mM Tris pH 7.5, 100 mM NaCl at -80 ° C.

In vitro Competitive Displacement Binding Assays for XIAP, cIAP-I and cIAP-2

The modified SMAC peptides and compounds were tested for their ability to replace the fluorescent tracer from XIAP, cIAP-1 or cIAP-2. The BIR3 region of cIAP-1, cIAP-2, and XIAP and the test compound or SMAC-based peptide and its corresponding peptide probe (Peptide Protein Research) in test buffer (50mM Hepes pH 7.5, 0.025% Tween-20, 0.01% BSA, and 1 mM DTT). The positive control group consists of the BIR3 protein and the tracker (no inhibition), while the negative control group contains only the tracker (100% inhibition). Incubate the samples at room temperature for 1 hour (XIAP and cIAP-2) or 3 hours (cIAP-1), and then use BMG Pherastar to analyze the chemical formula (Fluorescence Polarization mode) (FP 485nm, 520nm, 520nm, 520nm). IC ₅₀ values based (nonlinear least-squares analysis) by the use of a dose of nonlinear least squares analysis - plotting reaction determined.

Final conditions for XIAP, cIAP-1 and cIAP-2 tests

Anti-life-increasing

The cell growth inhibitory line uses the Alamar Blue test (Nociari, M.M., Shalev, A., Benias, P., Russo, C. Journal of Immunological Methods 1998, 213, 157-167). This method is based on the ability of living cells to reduce resazurin to its fluorescent product resorufin. For each proliferative assay, cells were plated in 96-well plates, the cells were allowed to recover for 16 hours, and then inhibitory compounds (at 0.1% DMSO v / v) were added for another 72 hours. At the end of the incubation period, 10% (v / v) Alamar Blue was added, followed by 6 hours of incubation, and fluorescence products were detected at 535 nM ex / 590 nM em.

The anti-proliferative properties of the compounds of the present invention are determined by measuring the ability of the compounds to inhibit the growth of three cancer cell lines:. EVSA-T (Human Breast Cancer) DSMZ cat. No. ACC 433. MDA-MB-231 (Human Breast Cancer) ECACC cat. No. 92020424. HCT116 (Human Colorectal Cancer) ECACC cat. No. 91091005 (non-sensitive cell line used as a control group in non-specific cytoxicity)

In tests using EVSA-T cell lines, the EC ₅₀ of Examples 1 to 34 was less than 0.01 μM. In particular, the EC ₅₀ values of Examples 1 to 3, 5 to 8, 10 to 14, 16, 18 to 25, 27 to 28, and 30 to 32 are less than 0.001 μM. In experiments using MDA-MB-231 cell lines, the EC ₅₀ of Examples 1 to 34 was less than 0.1 μM. In particular, the EC ₅₀ of Examples 1 to 8, 10 to 14, and 18 to 32 are less than 0.01 μM. More specifically, the EC ₅₀ of Examples 7 to 8 is less than 0.001 μM. These test data for the compounds of the present invention are shown in Table 1.

Apoptosis Induction

The following table summarizes the evaluation of the sensitivity to apoptosis induction of a group of 9 human melanoma cell lines, which were induced for 24 hours in the presence of 1 ng / ml TNF-α, 1 ng / ml TNF -α was added simultaneously with Example 2 of 1 µM. The range of sensitivity was observed with three cell lines (SK-MEL-24, WM-266-4, and WM-115) showing the least sensitivity (apoptosis <20% of cells after 24 hours). The table details the cytometry using a fluorescent caspase-3 matrix (NucView488-Biotium) after 24 hours of treatment with Example 2 at 1 µM plus 1 ng / ml TNF-α. ) To obtain the percentage of the total number of cells that are positive for lysing-cystease-3 activity.

HEK293-XIAP-Caspase-9 Immunoprecipitation (IP) MSD Test Method (Assay Protocol)

Stable HEK293-XIAP-Caspase-9 cells were plated in a 96-well plate [200 µl / well, 1 x 10 ⁶ cells / mL in cultured complete medium ( DMEM + 10% FBS + 0.5 mg / mL Geneticin (Invitrogen)], and restore overnight at 37 ° C. At 37 ° C, the compound in 0.1% DMSO will be added to the culture wells (duplicate) (duplicate) wells) for 2 hours. Place cells in 50 µl of 1 x MSD lysis buffer (1% Triton X-100 in 20 mM Tris.Cl (pH 7.6) and 150 mM NaCl containing protease inhibitor). Rocking at room temperature for 20 minutes to lysed. Streptavidin high binding MSD disc (L15SB-2) was coated with biotinylated anti-FLAG M2 antibody (Sigma F9291) in an amount of 25 µl / Well, diluted 5 µg / mL antibody in PBS, shake for 1 hour, and then block with 150 µl 3% BSA / TBST for 1 hour. Add cell lysate (25 µl) to the 96 wells The anti-FLAG coated MSD dish was placed on a shaker at room temperature for 4 hours. Washed with 150 µl TBST (20 mM Tris.Cl (pH 7.6), 150 mM NaCl, 0.1% Tween-20) 4 After Dilute with MSD blocking buffer (3% BSA / TBST) to 5 µl / mL of anti-Caspase-9 [CST # 9505] overnight at 4 ° C. Wash the discs at 150 µl TBST 4 After that, an anti-rabbit-sulfo tag (MSD cat no. R32AB-1) diluted to 2 µg / mL with MSD blocking buffer was added at room temperature for 2 hours. 150 µl of TBST Wash the disc 4 times and add 150 µl / well of 1 x MSD Interpretation Buffer (R92TC-2) before discriminating each disc.

EC ₅₀ values based (nonlinear least-squares analysis) by the use of a dose of nonlinear least squares analysis - plotting reaction determined. The EC ₅₀ of Examples 1 to 37 is less than 0.1 μM. In particular, the EC ₅₀ of Examples 1 to 13, 15, 18 to 25, 27 to 28, 30 to 34, and 36 to 37 are less than 0.01 μM. More specifically, the EC ₅₀ of Examples 10, 12, 23-24, and 31 is less than 0.001 μM. The test data of the compounds of the present invention are shown in Table 1.

Protocol for cIAP1 Degradation MSD Assay in MDA-MB-231 Cells

The MDA-MB-231 cells were placed in 96-well plate were plated (plating) [200 μl / hole, 4 x10 ⁵ cells / mL in complete medium (cultured complete medium) (DMEM + 10% FBS) in the culture] , And overnight at 37 ° C to recover. At 37 ° C, 0.1% DMSO compound was added to duplicate wells for 2 hours. The cells were rocked at room temperature in 50 µl of 1 x MSD lysis buffer (1% Triton X-100 in 20 mM Tris.Cl (pH 7.6), 150 mM NaCl containing a protease inhibitor). ) 20 minutes to lysed. Streptavidin high-binding MSD disk (L15SB-2) was coated with biotinylated anti-cIAP1 antibody (R & D Systems cat no. AF8181-internal biotinylated (biotinylated in house)) in an amount of 25 µl / The wells were diluted with 5 µg / mL antibody in PBS, shaken for 1 hour, and then blocked with 150 µl of 3% BSA / TBST for 1 hour. Cell lysate (25 µl) was added to the 96-well anti-cIAP1-coated MSD dish and placed overnight at 4 ° C. After washing 4 times with 150 µl TBST (20 mM Tris.Cl (pH 7.6), 150 mM NaCl, 0.1% Tween-20), add anti-dilution diluted to 6 µl / mL with MSD blocking buffer at room temperature. Anti-cIAP1-sulfo tag detection antibody (R & D Systems cat no. AF8181-internal sulfo-tagged in house) was over 2 hours. Wash the disc 4 times with 150 µl TBST and add 150 µl / well of 1 x MSD reading buffer (R92TC-2) before reading each dish.

IC ₅₀ value from the dose system using non-linear least squares analysis - was plotted to determine the reaction. The EC ₅₀ of Examples 1 to 37 is less than 0.01 μM. In particular, the EC ₅₀ values of Examples 1 to 8, 10 to 14, 16, 18 to 27, 30 to 34, and 37 are less than 0.001 μM. More specifically, the EC ₅₀ of Example 7 is less than 0.0001 μM. The test data of the compounds of the present invention are shown in Table 1.

Population patch clamp (PPC) assay protocol

The inhibition of hERG channels in CHO K1 cells was measured by an automated patch clamp test. These CHO K1 cell lines were stably transfected with hERG ion channels. PPC measurements were performed using an IonWorks Quattro instrument (Molecular Devices Corporation, Union City, CA) (which uses a 384-well PatchPlate (Molecular Devices Corporation) with 64 apertures per well). The concentration of the test compound is tested in the wells (two replicates). Amphotericin B was used to obtain electrical access to the inside of the cells, with a final concentration of 200 μg / mL. Measure human ether-à-gogo related gene (hERG) current using a prepulse from -80mV holding potential to +40 mV (2 s), then one step To -50 mV (2 s) to induce deactivating tail currents, then return to the holding potential for 1 second (1 s). Compounds were incubated for 600 seconds between pre- and post-compound reads. The external recording solution used was 130 mM sodium gluconate (Na Gluconate), 20 mM NaCl, 4 mM KCl, 1 mM MgCl ₂ , 1.8 mM CaCl ₂ , 10 mM Hepes, 5 mM glucose, and the pH was adjusted to 7.3 with NaOH. Filter all data for seal quality, seal drop, and current amplitude. Calculate the maximum current amplitude of the third pulse tail current before (Pre) and after (post) addition of the compound, and estimate the blocking amount by dividing the current amplitude after adding the compound by the current amplitude after adding the compound . The data generated using this test is detailed in Table 1.

Manual Patch Clamp (MPC) assay protocol

The inhibition of hERG channels in HEK293 cells was measured by a manual patch clamp test. These HEK293 cell lines were stably transfected with hERG ion channels. Use HEKA EPC10 amplifier and PatchMaster software to collect and analyze the data of this project.

Cells were placed on glass coverslips for plate culture, placed on an inverted microscope and continuously immersed in a control solution (137mM NaCl, 4mM KCl, 1mm MgCl ₂ , 1.8mM CaCl ₂ , 10mM Hepes, 10mM Glucose, pH 7.35).

After the cells were clamped electrically, they were left to balance and subjected to a pulse protocol. The pulse scheme involves stepping from a holding potential of -80mV to +40 mV for 4 seconds (4 s) to deactivate the hERG channel, and then stepping the membrane potential back to -50 mV for 4 seconds (4 s) ) To draw the tail current, and then return to the holding potential. This sequence is repeated at 20 second intervals between pulses. A voltage protocol was performed throughout the experiment. The experiment was started before administration (0.33% DMSO control group) and after cumulative addition of increasing compound concentrations. The evoked peak current amplitudes were continuously monitored throughout the experiment.

Before measuring the effect of the compound, first apply a potential to the test compound for 5 minutes or until the state is stable. Depending on which comes first, measure the spike tail current before and after the addition of each compound. The results of individual cells were normalized according to their respective vehicle control groups. Repeated measurement of each compound concentration. Use 0.1µM Cisapride as the reference inhibitor.

Table 1

Unless otherwise stated, data are the result of a single experiment. When more than one data point is obtained, the average value (ie, geometric mean or arithmetic mean) of these data points in the previous table is represented to two significant figures.

Combination Protocol for Apoptosis

One day before the treatment, melanoma cell lines (0.5 x 10 ⁶ cells / ml) were placed in the wells of a 24-well plate for plate culture (two replicates) to fixate them. After incubating the cells with the compounds for 24 hours in a CO ₂ incubator at 37 ° C with or without 1 ng / ml TNF-α (R & D Systems), the cells were harvested with trypsinisation. Cell pellets from 24-well plates were resuspended in 100 μl FACS buffer (PBS + 1% fetal bovine serum). Add NucView488 reagent (from Biotium) to a final concentration of 2 µM. After the plates were incubated in the dark for 30 minutes, the fluorescently stained cells were measured in a Guava easyCyte HT cytometer (Millipore). Lysing caspase-3 was recorded in the FL1 channel, and unstained and DMSO control groups were used to set gated stained or unstained cell populations.

Table 2 Summary Incubation for 24 hours in SK-MEL-28 or either A375 and any of the included agents shown in combination with Example 2 plus 1 ng / ml TNF-α, with% apoptosis observed Increase (the% apoptosis increases). When using the combination agents (with or without TNF-α) shown in the first column of the form alone, no increase in apoptosis was observed on this time scale and period-data not shown.

Table 2: Increased percentage of apoptotic cells after culturing using the combinations shown (relative to using only Example 2 + TNF-α *)

Combination Protocol for Cell Proliferation

The effect of the compound of the formula (I) (compound I) and the anticancer agent (compound II) can be evaluated by the following techniques. Cells from a human cell line (e.g., MDA-MB-231 and EVSA-T) respectively to 6.0 x10 ^3, a concentration of 2.5x10 ^3, or 4.0 x10 ³ cells / well, broadcasting (seeded) in 96-well tissue culture On the disk. Give cells 48 hours to recover before adding compounds or vehicle control (0.35% DMSO) as follows:

The compounds were added simultaneously for 96 hours. After a total of 96 hours of compound incubation, cells were fixed on ice for 1 hour using ice-cold 10% (w / v) trichloroacetic acid, and then plate washer (Labsystems Wellwash Ascent) at dH ₂ O Wash four times and air dry. Stain the cells with 0.4% (w / v) Sulforhodamine B (Sigma) (1% acetic acid) at room temperature for 20 minutes, then wash four times with 1% (v / v) acetic acid. Air dry and then add 10 mM Tris buffer to dissolve the dye. The colormetric product was read on a Wallac Victor ² plate reader (1420 multilabel counter, Perkin Elmer Life Sciences) at Abs 490nm for quantification. Measured at different doses of compound I compound II in the presence of the IC _50. Synergy was determined when the IC _{50 of} compound II migrated downward in the presence of sub-effective doses of compound I. When the reaction result of the compound I and the compound II together is equivalent to the total effect of the reactions of the two compounds individually, it is judged that there is additivity. The antagonistic effect IC ₅₀ defined as the upward migration, namely, the response to the two compounds is less than this sum of the individual effects of the two kinds of compounds.

no

[Figure 1] This is the ¹ H NMR of Example 39. The sample was obtained in DMSO-D ₆ and calibrated to the residual DMSO of the non-deuterium-substituted solvent as δ = 2.50 ppm. The internal reference standard (TCNB) contained a single peak. δ = 8.5 ppm. [Figure 2] XRPD of Example 39. [Figure 3] DSC of Example 39. [Figure 4] ¹ H NMR of Example 40; the sample was obtained in DMSO-D ₆ and calibrated to a non-deuterium-substituted solvent residual DMSO of δ = 2.50 ppm; the included internal reference standard (TCNB) showed a single peak δ = 8.5 ppm. [Figure 5] XRPD of Example 40. [Figure 6] DSC of Example 40. [Figure 7] This is the ¹ H NMR of Example 41; the sample was obtained in DMSO-D ₆ and calibrated to the residual DMSO of the non-deuterium-substituted solvent as δ = 2.50 ppm. [Fig. 8] XRPD of Example 41. [Figure 9] DSC of Example 41. [Figure 10] This is the ¹ H NMR of Example 42; the sample was obtained in DMSO-D ₆ and calibrated to the residual DMSO of the non-deuterium-substituted solvent as δ = 2.50 ppm. [Figure 11] XRPD of Example 42. [Figure 12] DSC of Example 42. [Figure 13] XRPD, which shows the product of L-(+)-lactate form B (diffraction icon 1) of Example 40, the reaction mixture (diffraction icon 2) at t = 0h, and 4 days later ( Diffraction icon shows 3), compared with L-(+)-lactate form C of Example 43 (diffraction icon shows 4). [Figure 14] XRPD, which shows the same iso-structural Form B product of Example 43, at t = 0h (diffraction icon shown 1), the progress of the reaction mixture (diffraction icon shown 2-6), heating After t = 5 days, the internal conversion is completed to obtain Form C (diffraction icon shows 7). [Figure 15] This is the ¹ H NMR of Example 43. The sample was obtained in DMSO-D ₆ and calibrated to the residual DMSO of the non-deuterium-substituted solvent as δ = 2.50 ppm. The included internal reference standard (TCNB) showed a single peak δ = 8.5 ppm. [Fig. 16] It is the XRPD of Example 43, (diffraction icon is shown as 1) and anhydrous L-(+)-lactic acid (diffraction icon is shown as 2) is overlapped. [Fig. 17] DSC of Example 43, (Thermal analysis icon 1) and the anhydrous L-(+)-lactic acid (Thermo analysis icon 2) are overlapped.

Claims (61)

A compound, as shown in formula (I):

Or a tautomer thereof, a stereochemical isomer thereof, a pharmaceutically acceptable salt thereof, or a solvate thereof; wherein X is CR ⁴ , N or NR ³ ; wherein when X is CR ⁴ , then U Represents nitrogen and R ⁶ represents a side oxygen (oxo); or when X is N, U represents carbon and R ⁶ represents methylol or -CH (OR ^x ) CH ₂ OR ^z ; or when X is NR ³ , then U represents carbon and R ⁶ represents a side oxygen group; a dashed bond (-------) represents a single or double bond, wherein at least two of the dashed bonds represent a double bond; R ¹ and R ² is independently hydrogen or methyl; R ³ is hydrogen, methyl or -NH ₂ ; R ⁴ is hydrogen, methyl, methylol, -NH ₂ or fluorine; R ⁵ is unsubstituted n-butyl, or one or two of fluoro-substituted benzyl group on the phenyl; and R ^x represent, and R ^z independently represent hydrogen or methyl lines. The compound according to item 1 of the scope of patent application, wherein one of R ¹ and R ² represents hydrogen and the other represents methyl, or both R ¹ and R ² represent hydrogen. The compound as described in item 1 of the scope of patent application, wherein both R ¹ and R ² represent hydrogen. The compound as described in claim 1 in the scope of patent application, wherein R ⁴ represents hydrogen or methyl. The compound as described in item 1 of the scope of patent application, wherein R ⁵ represents unsubstituted n-butyl or benzyl substituted with one or two fluorines at positions 2, 3, and / or 4 of phenyl. The compound as described in claim 5 in the scope of patent application, wherein R ⁵ represents unsubstituted n-butyl. The compound as described in claim 5 in the scope of the patent application, wherein R ⁵ represents a benzyl group substituted with a fluorine at position 2, 3 or 4 of the phenyl group. The compound as described in item 5 of the scope of patent application, wherein R ⁵ represents 2-fluorobenzyl, 3-fluorobenzyl or 4-fluorobenzyl. The compound as described in item 5 of the scope of patent application, wherein R ⁵ represents a benzyl group substituted with a fluorine at position 4 of the phenyl group. The compound as described in item 5 of the scope of patent application, wherein R ⁵ represents a benzyl group substituted with two fluorines at positions 2 and 3, 3 and 4, or 2 and 4 of the phenyl group. The compound as described in item 5 of the scope of patent application, wherein R ⁵ represents 2,3-difluorobenzyl, 3,4-difluorobenzyl, or 2,4-difluorobenzyl. The compound as described in item 5 of the scope of patent application, wherein R ⁵ represents a benzyl group having two fluorine substitutions at positions 2 and 4 of the phenyl group. The compound as described in item 5 of the scope of patent application, wherein R ⁵ represents 4-fluorobenzyl or 2,4-difluorobenzyl. The compound as described in claim 13 in which R ⁵ represents 2,4-difluorobenzyl. The compound as described in claim 13 in the scope of patent application, wherein R ⁵ represents 4-fluorobenzyl. The application of the compound of item 1 patentable scope, wherein one of R ^x and R ^z are hydrogen and the other represents methyl, or both R ^x and R ^z are both hydrogen. The compound as described in claim 16 in the scope of patent application, wherein R ^x represents hydrogen or methyl, and R ^z represents hydrogen. The compound as described in item 1 of the scope of patent application, which is a compound of formula (Ia):

Or tautomers thereof, stereochemical isomers thereof, pharmaceutically acceptable salts thereof, or solvates thereof; wherein R ¹ , R ² , R ⁴ and R ^{5 are} as described in items 1 to 15 of the scope of patent application Any one of them. The compound as described in item 1 of the scope of patent application, which is a compound of formula (Ib):

Or a tautomer thereof, a stereochemical isomer thereof, a pharmaceutically acceptable salt thereof, or a solvate thereof; wherein R ⁶ represents methylol or -CH (OR ^x ) CH ₂ OR ^z , and R thereof ¹ , R ² , R ⁵ , R ^x and R ^{z are} as described in any one of claims 1 to 3 and 5 to 17 of the patent application scope. The compound as described in item 1 of the scope of patent application, which is a compound of formula (Ic):

Or tautomers thereof, stereochemical isomers thereof, pharmaceutically acceptable salts thereof, or solvates thereof; wherein R ¹ , R ² , R ³ and R ^{5 are} as in the scope of application for patents Nos. 1 to 3 and 5 As described in any one of 15 to 15. The compound as described in item 1 of the scope of patent application, which is a compound of formula (Id):

Or a tautomer thereof, a stereochemical isomer thereof, a pharmaceutically acceptable salt thereof, or a solvate thereof; wherein R ^{5 is} as described in any one of claims 1 and 5 to 15 of the patent application scope. The compound as described in item 1 of the scope of patent application, wherein the compound is selected from Examples 1 to 37:

Or a tautomer thereof, a stereochemical isomer thereof, a pharmaceutically acceptable salt thereof, or a solvate thereof. The compound as described in item 1 of the scope of patent application, wherein the compound is Example 2:

Free base, or a tautomer thereof, a stereochemical isomer thereof, a pharmaceutically acceptable salt thereof, or a solvate thereof. The compound according to item 1 of the scope of patent application, wherein the compound is 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H , 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholine- 4-yl] methyl} piperazin-1-yl] ethan-1-one is a lactate, mesylate, or sulfate. The compound according to item 1 of the scope of patent application, wherein the compound is 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H , 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholine- L-(+)-lactate of 4-yl] methyl} piperazin-1-yl] ethan-1-one. The compound according to item 1 of the scope of patent application, wherein the compound is 1- {6-[(4-fluorophenyl) methyl] -5- (hydroxymethyl) -3,3-dimethyl-1H , 2H, 3H-pyrrolo [3,2-b] pyridin-1-yl} -2-[(2R, 5R) -5-methyl-2-{[((3R) -3-methylmorpholine- 4-yl] methyl} piperazin-1-yl] ethan-1-one L-(+)-lactate (type C). A pharmaceutical composition comprising the compound of the formula (I) as described in any one of claims 1 to 26 and a pharmaceutically acceptable excipient. A pharmaceutical composition comprising a compound of formula (I) as described in any one of claims 1 to 26 of the scope of patent application, and one or more therapeutic agents. The compound according to any one of claims 1 to 26 of the scope of patent application, which is used for treatment. The compound according to any one of claims 1 to 26 of the scope of patent application, which is used to treat a disease state or abnormal physical condition caused by IAP. The compound according to any one of claims 1 to 26 of the scope of patent application, which is used for treating a disease state or abnormal physical condition caused by XIAP and / or cIAP. The compound according to any one of claims 1 to 26 of the scope of patent application, which is used to treat a disease state or abnormal physical condition of IAP overexpression. The compound according to any one of claims 1 to 26 of the scope of patent application, which is used for treating a disease state or abnormal physical condition of XIAP and / or cIAP over-expression. A use of a compound according to any one of claims 1 to 26 of the scope of patent application for the preparation of a medicament for treating cancer. The use according to item 34 of the application, wherein the cancer is an epithelial tumor; a hematological malignancy and a precancerous blood disease and a borderline malignancy; a mesenchymal tumor; a tumor of the central or peripheral nervous system; Ocular and ocular tumors; germ cell and trophoblastoma; children and embryo tumors. The use according to item 34 of the application, wherein the cancer is the bladder and urethra, breast, gastrointestinal tract, liver, gallbladder and bile duct system, exocrine pancreas, kidney, lung, head and neck, ovary, fallopian tube, peritoneal Malignant tumors of the vagina, vulva, penis, cervix, myometrium, endometrium, thyroid, adrenal gland, prostate, skin or skin. The use according to item 36 of the application, wherein the malignant tumor of the lung is adenocarcinoma, small cell lung cancer, non-small cell lung cancer, bronchioloalveolar carcinoma or mesothelioma. The use as described in claim 34, wherein the cancer is mesothelioma. The use according to item 34 of the application, wherein the cancer is liver cancer, melanoma, esophageal cancer, kidney cancer, colon cancer, rectal cancer, lung cancer, breast cancer, bladder cancer, gastric cancer, ovarian cancer or prostate cancer. The use according to item 34 of the application, wherein the cancer is kidney cancer, melanoma, colon cancer, lung cancer, breast cancer, ovarian cancer or prostate cancer. The use according to item 34 of the application, wherein the cancer is melanoma, colon cancer, breast cancer or ovarian cancer. The use according to item 34 of the scope of patent application, wherein the cancer is melanoma. The use according to item 34 of the scope of patent application, wherein the cancer is breast cancer. The use according to item 34 of the scope of patent application, wherein the cancer is inflammatory breast cancer. The use according to item 34 of the scope of patent application, wherein the cancer is an inflammatory tumor. The use according to item 45 of the scope of patent application, wherein the inflammatory tumor is melanoma, colon cancer, breast cancer or ovarian cancer. The use according to item 45 of the scope of patent application, wherein the inflammatory tumor is melanoma. The use as described in claim 34, wherein the cancer is a hematological malignancy. The use as described in claim 34, wherein the cancer is leukemia or lymphoma. The use according to item 49 of the scope of patent application, wherein the lymphoma is a MALT lymphoma. The use according to item 34 of the application, wherein the cancer is selected from the group consisting of acute lymphoblastic leukemia (ALL), chronic lymphocytic leukemia (CLL), B-cell lymphoma, follicular lymphoma, Burkitt's lymphoma, Mantle cell lymphoma, T cell lymphoma and leukemia, natural killer cell (NK) lymphoma, Hodgkin's lymphoma, hair cell leukemia, unknown monoclonal gammopathy, plasma cell tumor, multiple bone marrow Tumor and lymphoproliferative disorders after transplantation. The use according to item 51 of the scope of patent application, wherein the B-cell lymphoma is diffuse large B-cell lymphoma (DLBCL). The use according to item 51 of the scope of patent application, wherein the B-cell lymphoma is refractory DLBCL. The use as described in claim 34, wherein the cancer is selected from the group consisting of acute myeloid leukemia (AML), chronic myelogenous leukemia (CML), chronic bone marrow mononuclear leukemia (CMML), and eosinophilia Disease (hypereosinophilic syndrome), myeloproliferative disorders (myeloproliferative disorders), myeloproliferative syndrome, myelodysplastic syndrome and promyelocytic leukemia. The compound according to any one of claims 1 to 26 of the scope of patent application, which is: one or more other therapeutic agents; or one or two other therapeutic agents; or one or more other anticancer agents; or 1 Or a combination of 2 other anticancer agents. The compound according to any one of claims 1 to 26 of the scope of patent application, which is: one or more other therapeutic agents; or one or two other therapeutic agents; or one or more other anticancer agents; or 1 Or 2 other anticancer agents for treatment. The compound according to any one of claims 1 to 26 of the scope of patent application, which is: one or more other therapeutic agents; or one or two other therapeutic agents; or one or more other anticancer agents; or 1 Or a combination of 2 other anticancer agents for treating cancer. A method for preparing a compound of formula (I) as claimed in item 1 of the patent application scope, comprising: (a) (i) combining a compound of formula (II)

Among them, R ⁵ , R ⁶ , U, and X are as described in the compound of formula (I) in the first patent application, L ¹ represents a suitable leaving group, and P ¹ represents hydrogen or a suitable protecting group. Reacts with a compound of formula (III):

Or a selectively protected derivative thereof; wherein R ¹ and R ^{2 are} as described in the compound of formula (I) in the first patent application range, and then suitable for removing the P ¹ protecting group, and, if necessary, any Other protecting groups, the deprotection reaction; or (ii) the compound of formula (IV):

Among them, R ⁵ , R ⁶ , X and U are as described in the compound of formula (I) in the first patent application range, and L ² represents a suitable leaving group to react with the compound of formula (V):

Or its selectively protected derivative; wherein R ¹ and R ^{2 are} as described in the compound of formula (I) in the first patent application, and P ² represents hydrogen or a suitable protecting group, and then suitable removal of P is performed. ² protecting group, and, if necessary, any other protecting group, a deprotection reaction; and / or (b) a deprotection reaction of a protected derivative of the compound of formula (I); and / or (c) I) Interconversion of a compound or a protected derivative thereof into another compound of formula (I) or a protected derivative thereof; and (d) a pharmaceutically acceptable compound that selectively forms a compound of formula (I) salt. The method as described in claim 58 in which L ¹ and L ² represent halogen atoms. The method according to item 58 of the scope of patent application, wherein L ¹ represents chlorine. The method according to item 58 of the scope of patent application, wherein P ¹ and P ² represent a third butoxycarbonyl group (tBoc).